2W Groups Research Articles

(057) Influence of Circadian Disruption on Erectile Function in Chronic Jet-Lagged Young Rats

Abstract Introduction Sleep is crucial during the developing stage of one’s life and has been associated with physical activity. However, as the number of 24-hour stores such as convenience stores and supermarkets have increased, along with increased internet usage, people have developed the habit of staying up late, resulting in sleep deprivation due to disruption in the light-dark cycle. Jet lag is widely used in animal models for inducing irregular circadian rhythms. Chronic jet lagged model animals are known to develop a variety of disorders. Objective To investigate the influences of chronic jet lag on erectile function in rats in order to elucidate the effects of irregular circadian rhythms on future sexual function. Methods 4-week-old rats underwent randomly assigned to remain either under the control light/dark cycle (Cont group) or under chronic light/dark shift schedule to simulate the conditions of chronic jet lag (CJL group). The shift-lag protocol consisted of 8 h light/dark phase advances repeated every 3 days for 4 weeks. Erectile function was examined at 1, 2, and 4 weeks post CJL treatment. After 4 weeks of CJL treatment, the rats were returned to control light/dark cycle for 4 and 8 weeks, as CJL(4w)+Rest(4w) group and CJL(4w)+Rest(8w) group, and erectile function was examined correspondingly. Erectile function was tested using intracavernous pharmacotherapy (ICP) measurements, and endothelial function was tested through an isometric tension study. Western blot analysis was used to assess the levels of endothelial nitric oxide (eNOS) and phosphorylated eNOS (p-eNOS). Additionally, mRNA expressions of clock genes (Bmal1 and Cry 1) and growth factors (TGF-β1 and ICAM-1) were tested using real-time PCR. Results The ICP/MAP ratio was significantly lower in the CJL group (0.35 ± 0.05) at 4 weeks than in the control group (0.66 ± 0.05) (P < 0.05). CJL group exhibited significantly lower testosterone levels, Bmal1, Cry 1, TGF-β1, and ICAM-1 expression, and p-eNOS and cGMP levels. The ICP/MAP ratio in the CJL(4w)+Rest(4w) group (0.47 ± 0.06) was lower than that in the control group (P < 0.01) whereas the ratio in the CJL(4w)+Rest(8w) group (0.81 ± 0.03) was similar to that in the control group (P > 0.05). Conclusions In this study, chronic jet lagged (4 weeks) rats exhibited a decreased ICP/MAP ratio, reduced testosterone level, and decreased p-eNOS expression, indicating the development of erectile dysfunction (ED). Upon 8 weeks of normal light/dark shift schedule, erectile function was recovered. This study suggests that irregular circadian rhythms induced by repeated jet lag, especially in childhood, may affect the production of sex hormones and erectile function in the future. Therefore, it is necessary for children to follow a regular circadian rhythm and healthy lifestyle. Disclosure No.

Evaluation of the effect of diode laser application on the hydrophilicity, surface topography, and chemical composition of titanium dental implant surface.

Attempts have been made to decontaminate the implant surface by using diode lasers. However, the parameters that provide efficient decontamination without altering the topography or surface characteristics of titanium implants are still unclear. The aim of the present study was to evaluate the effect of altering the power of diode laser (940nm) application on the hydrophilicity, surface topography, and chemical composition of sandblasted, large grit, acid-etched (SLA) treated titanium alloy implant surface. Thirty-six SLA-treated titanium discs (Dentis Co., Ltd.) were used in this study. The hydrophilicity of all discs was measured by using a contact angle goniometer (190 CA; Rame-hart Co, Ltd). Discs were randomly divided into four groups (n = 9 each) based on the power of the diode laser used. Group I (control, no lasing group), group II: treated with 1W power, group III: 2W power, and group IV: 3W power. The chemical composition of the SLA discs was evaluated by using energy dispersive x-ray spectroscopy (EDX) before laser application. Hydrophilicity was reevaluated after the application of laser irradiation. The surface topography of all discs was examined. Changes in the chemical composition of the titanium discs were investigated following the lasing procedure. Morphometric analysis of the surface area (μm2 ) of the indentations created following laser application was also evaluated. Data were collected and the Shapiro-Wilk test of normality was used. Comparisons between the four study groups were done by using the Kruskal-Wallis test, while that to evaluate the morphometric analysis of the surface area was done by using One-way ANOVA (P<0.05). The average contact angle of the drop of distilled water to the SLA discs significantly decreased after laser treatment (P<0.05). The largest contact angle was measured in the control group, followed by the 1W group, and the 2W group. The smallest angle was measured in the 3W group. Considerable surface alterations such as melting and flattening were observed on examination of the surface topography of the 3W group followed by the 2W group. The least changes were observed in association with the 1W group in comparison to the control group. The EDX analysis showed the appearance of peaks of the oxygen and carbon elements after the lasing procedure with the highest percentage in the 3W group. The average of the surface area of the created indentations significantly increased with increasing the power of the diode laser used (P<0.05). The application of diode laser (940nm) with 2W and 3W powers significantly altered the hydrophilicity, the surface topography, and the chemical composition of titanium discs. Diode laser (940nm) with 1W power can be safely used on SLA titanium implant surfaces with no damaging effect on the surface topography or hydrophilicity.

Role of osteopontin in the process of pulpal healing following tooth replantation in mice.

IntroductionThe role of osteopontin (OPN) following severe injury remains to be elucidated, especially its relationship with type I collagen (encoded by the Col1a1 gene) secretion by newly-differentiated odontoblast-like cells (OBLCs). In this study, we examined the role of OPN in the process of reparative dentin formation with a focus on reinnervation and revascularization after tooth replantation in Opn knockout (KO) and wild-type (WT) mice.MethodsMaxillary first molars of 2- and 3-week-old-Opn KO and WT mice (Opn KO 2W, Opn KO 3W, WT 2W, and WT 3W groups) were replanted, followed by fixation 3–56 days after operation. Following micro-computed tomography analysis, the decalcified samples were processed for immunohistochemistry for Ki67, Nestin, PGP 9.5, and CD31 and in situ hybridization for Col1a1.ResultsAn intense inflammatory reaction occurred to disrupt pulpal healing in the replanted teeth of the Opn KO 3W group, whereas dental pulp achieved healing in the Opn KO 2W and WT groups. The tertiary dentin in the Opn KO 3W group was significantly decreased in area compared with the Opn KO 2W and WT groups, with a significantly low percentage of Nestin-positive, newly-differentiated OBLCs during postoperative days 7–14. In the Opn KO 3W group, the blood vessels were significantly decreased in area and pulp healing was disturbed with a failure of pulpal revascularization and reinnervation.ConclusionsOPN is necessary for proper reinnervation and revascularization to deposit reparative dentin following severe injury within the dental pulp of erupted teeth with advanced root development.

Expansion of the Inguinal Adipose Tissue Depot Correlates With Systemic Insulin Resistance in C57BL/6J Mice.

To accommodate surplus energy, the adipose tissue expands by increasing adipocyte size (hypertrophy) and number (hyperplasia). The presence of hypertrophic adipocytes is a key characteristic of adipose tissue dysfunction. High-fat diet (HFD) fed C57BL/6J mice are a commonly used model to study obesity and obesity-related complications. In the present study, we have characterized adipose plasticity, at both the cellular and tissue level, by examining the temporal development of systemic insulin resistance and adiposity in response to HFD-feeding for 4, 8, and 12 weeks (4w, 8w, and 12w). Within the same time frame, we examined systemic metabolic flexibility and adipose plasticity when switching from HFD- to chow-diet during the last 2 weeks of diet intervention (referred to as the reverse (REV) group: 4wREV (2w HFD+2w chow), 8wREV (6w HFD+2w chow), 12wREV (10w HFD+2w chow)). In response to HFD-feeding over time, the 12w group had impaired systemic insulin sensitivity compared to both the 4w and 8w groups, accompanied by an increase in hypertrophic inguinal adipocytes and liver triglycerides. After reversing from HFD- to chow-feeding, most parameters were completely restored to chow control levels for 4wREV and 8wREV groups. In contrast, the 12wREV group had a significantly increased number of hypertrophic adipocytes, liver triglycerides accumulation, and impaired systemic insulin sensitivity compared to chow-fed mice. Further, image analysis at the single-cell level revealed a cell-size dependent organization of actin filaments for all feeding conditions. Indeed, the impaired adipocyte size plasticity in the 12wREV group was accompanied by increased actin filamentation and reduced insulin-stimulated glucose uptake compared with chow-fed mice. In summary, these results demonstrate that the C57BL/6J HFD-feeding model has a large capacity to restore adipocyte cell size and systemic insulin sensitivity, and that a metabolic tipping point occurs between 8 and 12w of HFD-feeding where this plasticity deteriorates. We believe these findings provide substantial understanding of C57BL/6J mice as an obesity model, and that an increased pool of hypertrophic ING adipocytes could contribute to aggravated insulin resistance.

Maintenance of increased bone mass after PTH withdrawal by sequential medicarpin treatment via augmentation of cAMP-PKA pathway.

Osteoporosis is a metabolic bone disorder associated with impaired bone microarchitecture leading to fragility fractures. Long-term usage of parathyroid hormone (PTH) enhances bone resorption and leads to osteosarcoma in rats which limits its exposure to maximum 2 years in human. Notably, the anabolic effects of PTH do not endure in the absence of sustained administration. Studies in our lab identified osteogenic and antiresorptive activity in medicarpin, a phytoestrogen belonging to the pterocarpan class. Considering dual-acting property of medicarpin and limitations of PTH therapy, we envisaged that medicarpin sequential treatment after PTH withdrawal could serve as promising therapeutic approach for osteoporosis treatment. As PTH exerts its bone anabolic effect by increasing osteoblast survival, our study aims to determine whether medicarpin amplifies this effect of PTH. Our results show that PTH withdrawal led to reduced bone mineral density and bone parameters, while sequential treatment of medicarpin after PTH withdrawal significantly enhanced these parameters. Remarkably, these effects were more pronounced than 8-week PTH treatment. Sequential therapy also significantly increased P1NP levels and decreased CTX levels and TRAP positive cells compared to PTH 8W group where CTX levels were quite high due to bone resorptive action of PTH. Protein expression studies revealed that medicarpin along with PTH betters the antiapoptotic potential compared to PTH alone, through augmentation of cyclic adenosine monophosphate-PKA-CREB pathway. These results proclaim that medicarpin sequential treatment prevented the reduction in bone accrual and strength accompanying PTH withdrawal and also aided in antiapoptotic role of PTH. The study points toward the potential use of medicarpin as a replacement therapeutic option postdiscontinuation of PTH.

Irisin/FNDC5 influences myogenic markers on skeletal muscle following high and moderate-intensity exercise training in STZ-diabetic rats.

In the present study, we investigated the effects of high-intensity interval training (HIIT) versus moderate-intensity continuous training (MICT) on irisin and expression of myogenic markers (paired box 7 (Pax7), myogenic differentiation 1 (MyoD), myogenin) in skeletal muscle of diabetic rats. Eighty-four male Wistar rats (6weeks of age) were randomly divided into seven groups (n = 12): basic control (Co Basic), 8weeks control (Co 8w), diabetes mellitus (DM), HIIT, DM + HIIT, MICT, and DM + MICT groups. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ). The protocol was characterized by running on a rodent treadmill with moderate intensity (60-70% ), 60min per session, 5days/week, for 6weeks. HIIT consisted of six 3-min runs at a high intensity (80-90% ) alternated with 2-min running at low intensity (50% ), 30min per session, 5days/week, for 6weeks. Results showed that DM decreased myoblast markers compared to Co Basic and Co 8w groups. Fibronectin type III domain-containing protein 5 (FNDC5) mRNA decrease was correlated with myoblast markers (Pax7 r = 0.632, p = 0.027; MyoD r = 0.999, p = 0.001; myogenin r = 1.000, p = 0.001) in DM group. DM + MICT significantly increased gene expression of MyoD, myogenin, and FNDC5 compared to DM and DM + HIIT. The results also showed that the intensity and duration of exercise on the treadmill were effective in stimulating irisin and myogenic markers after DM.

The potential role of herbal extract Wedelolactone for treating particle-induced osteolysis: an in vivo study

BackgroundOsteolysis is one of the most prevalent clinical complications affecting people who undergo total joint replacement (TJR). Wedelolactone (WDL) is a coumestan compound derived from the Wedelia chinensis plant and has been demonstrated to exhibit anti-inflammatory properties. This study aimed to investigate the oral administration of WDL as a potential treatment for particle-induced osteolysis using a well-established mice calvarial disease model.MethodsThirty-two C57BL/6 J mice were randomized into four groups: Sham, vehicle, osteolysis group with oral WDL treatment for 4 weeks (WDL 4w), and osteolysis group treated for 8 weeks (WDL 8w). Micro-CT was used to quantitatively analyze the bone mineral density (BMD), bone volume/tissue volume (BV/TV) and trabecular bone thickness (Tb.Th). Osteoclast numbers were also measured from histological slides by two investigators who were blind to the treatment used.ResultsThe results from micro-CT observation showed that BMD in the WDL 8w group improved significantly over the vehicle group (p < 0.05), but there was no significant difference between WDL 4w and 8w for BV/TV and Tb.Th. Osteoclast numbers in the WDL 4w group were also lower than the vehicle group (p < 0.05), but the difference between WDL 8w and 4w groups was not significant.ConclusionsParticle-induced osteolysis is an inevitable long-term complication after TJR. The results of this animal study indicate that an oral administration of WDL can help reduce the severity of osteolysis without adverse effects.

Identification of key genes associated with the progression of liver fibrosis to hepatocellular carcinoma based on iTRAQ proteomics and GEO database

Introduction and objectiveLiver fibrosis (LF) often leads to cirrhosis and even hepatocellular carcinoma (HCC), but the molecular mechanism remains unclear. The aims of the present study were to identify potential biomarkers for the progression of LF to HCC and explore the associated molecular mechanisms. Materials and methodsThe isobaric tags for relative and absolute quantitation (iTRAQ) was used to detect changes in the protein expression profiles of liver tissues and to screen the differentially expressed proteins (DEPs). The differentially expressed genes (DEGs) of LF rats and patients were screened by Gene Expression Database (GEO). Subsequently, the clinicopathological analysis of the overlapping genes in different pathological stages in HCC patients based on GEPIA database was conducted. ResultsiTRAQ proteomic analysis revealed 689, 749 and 585 DEPs in the 6W, 8W and 12W groups, respectively. ALDH2, SLC27A5 and ASNS were not only the DEPs found in rats with LF with different stages but were also the DEGs related to the pathological stages and survival in patients with HCC. ConclusionsALDH2, SLC27A5 and ASNS were the potential biomarkers associated with the progression of LF to HCC.

How to Establish a Minimal Invasive and Stable Carotid Artery Stenosis Rabbit Model? A Simple and Effective Carotid Artery Balloon Strain Technique.

Background: The objective of this study is to establish a minimally invasive technique to create a stable carotid artery stenosis rabbit model. This article summarizes the specific methods and key points of this technology.Methods: The experiment studied a rabbit that was anesthetized through the vein. After the femoral artery was exposed, a minimally invasive needle was used to puncture the femoral artery, then the sheath was placed into the artery. We primarily put a catheter in the ascending aorta for angiography and then used a PT2 guidewire for super-selection. The PT2 guidewire was retained, and a balloon was placed in the right common carotid artery (CCA) through a guidewire to inflate it three times. Six rabbits in the 2- (2W) and 4-week (4W) groups were examined at 14 and 28 days, respectively. The rabbits in the control group received angiography at the beginning and 28 days later but without balloon injury. After angiography assessment, specimens of right CCA were dissected. Pathological and immunohistochemical examinations were performed on the collected specimens, and iFlow analysis was performed as well.Results: All the 18 animals which survived were observed. The rabbits in the 2W and 4W groups showed stenosis of the right CCA. Digital subtraction angiography showed the diameter was lower than that in the control group (1.04 ± 0.1, 0.71 ± 0.12, and 1.83 ± 0.08 mm in 2W, 4W, and control group, P < 0.05). Pathology also suggested carotid stenosis and obvious intimal hyperplasia. The results of immunohistochemistry showed that α-smooth muscle actin was highly expressed in the 2W and 4W groups, and the integrated optical density (IOD) value was higher than that in the control group (14,807.11 ± 1,822.3, 22,245.96 ± 1,212.82, and 6,537.16 ± 1,186.62 in the 2W, 4W, and control group, P < 0.05). Meanwhile, a cluster of differentiation 31 (CD31) was low expressed in the 2W and 4W groups, and the IOD value was lower than that in the control group (519.14 ± 44.4, 1,029.64 ± 98.48, and 1,502.05 ± 88.79 in the 2W, 4W, and control group, P < 0.05), which suggested endothelial damage and partial repair. The analysis by iFlow showed that the time-to-peak after balloon strain in the 2W and 4W groups were longer than that in the control group.Conclusion: We established a minimally invasive, effective, and safe method to establish a carotid artery stenosis rabbit model. The highlights of this technology were the application of minimally invasive methods, reducing surgical bleeding, infection, and related complications. This technology avoided the influence of tissue around CCA in the traditional carotid artery balloon injury model, which might lead to more accurate treatment outcomes.

The protective mechanism of SIRT1 on cartilage through regulation of LEF-1

BackgroundOsteoarthritis (OA) is a chronic degenerative disease that suppresses middle-aged and older people worldwide. Silent information regulator 1(SIRT-1) is associated with several age-related diseases, such as cardiovascular diseases, neurodegenerative diseases and tumors, etc. The protective role of SIRT-1 in bone and joint diseases has become increasingly well known.ObjectiveTo explore the relationship between SIRT-1 and its related factors in OA.MethodsFresh tibial plateau specimens were collected from 30 patients with knee OA who underwent total knee arthroplasty. According to the results of Safranin O Fast Green Staining, hematoxylin–eosin staining and the OARSI grade developed by the International Association for the Study of Osteoarthropathy, the specimens were divided into the mild group, moderate group and severe group, and the damage of cartilage was evaluated. SIRT-1 protein levels in cartilage samples were analyzed by immunohistochemistry. Then, take 60 8-week-old female C57BL/6 J mice and apply the Destabilization of the medial meniscus (DMM) to induce OA. Mice were randomly divided into normal group (sham), model group (model), and post-modeling drug administration group (srt), and each group was further divided into 2 weeks after modeling (2 W) and 8 weeks after modeling (8 W) according to the time after surgery. The degenerative degree of a knee joint in mouse knee cartilage samples was evaluated using Safranin O Fast Green Staining and OARSI grade. Immunohistochemical techniques assessed the protein levels of SIRT-1, β-catenin, LEF-1, MMP-13 and Collagen II in cartilage samples. The protein levels of β-catenin, LEF-1 and MMP-13 in the samples were assessed by the immunohistofluorescence technique. The mRNA expression of SIRT-1 and LEF-1 in mouse cartilage samples was evaluated by real-time quantitative polymerase chain reaction (qPCR).ResultsIn the human cartilage samples, according to the results of Safranin O Fast Green Staining, compared with the mild group, the moderate group and the severe group showed damage cartilage layer structure, the number of chondrocytes decreased, the cell hypertrophic, the cartilage surface discontinuous, and the OARSI grade increased. The severe group had severe cartilage injury and the highest OARSI grade. In the mice cartilage samples, according to immunohistochemical analysis, the protein levels of β-catenin, LEF-1 and MMP-13 in cartilage specimens of model 2 W and model 8 W groups were significantly increased than the sham 2 W and sham 8 W groups. The protein levels of SIRT-1 and Collagen II were significantly decreased (P < 0.05), the results of srt 2 W and srt 8 W groups were between the sham group and the model group. According to immunofluorescence analysis, the protein levels of β-catenin, LEF-1 and MMP-13 in model 2 W and model 8 W groups were significantly increased than sham 2 W and sham 8 W groups. The results of srt 2w and srt 8w groups were between the sham group and the model group. According to the real-time qPCR results: Compared with sham 2 W and sham 8 W groups, the mRNA expression of SIRT-1 in model 2 W and model 8 W groups was significantly decreased, while the mRNA expression of LEF-1 was significantly increased. In contrast, the results of srt 2 W and srt 8 W groups were between the sham group and the model group.ConclusionSRT-1720, as a specific activator of SIRT-1, does increase the protein level of SIRT-1. SIRT-1 may play a protective role in cartilage by regulating the expression of LEF-1 and related inflammatory factors in OA.

Effect of Late Androgen Replacement Therapy on Erectile Function Through Structural Changes in Castrated Rats.

BackgroundBecause androgen replacement therapy (ART) is not performed immediately after the onset of androgen deficiency, the treatment is considered to be late.AimTo investigate the effects of late ART, starting 4 weeks after castration of rats, on erectile function and structural changes in the corpus cavernosum.MethodsRats were subjected to ART for 4 (Late-ART [4w]) or 8 (Late-ART [8w]) weeks. In either case, rats were assigned to the following groups: castrated (Cast), castrated with subcutaneous administration of testosterone (3 mg/kg/day; Cast+T), and sham (Sham). Cast + T rats received daily subcutaneous doses of testosterone starting 4 weeks after castration for 4 or 8 weeks whereas Sham and Cast rats received only the vehicle.OutcomesErectile function was assessed by evaluating intracavernosal pressure (ICP) and mean arterial pressure (MAP) after electrical stimulation of the cavernous nerve, corporal veno-occlusive function using dynamic infusion cavernosometry, and histology using Masson's trichrome staining.ResultsNo increase in the ICP was observed in Cast+T rats in the Late-ART (4w) group (0.47 ± 0.02, P > .05), whereas, in Cast+T rats in the Late-ART (8w) group, there was a significant increase in the ICP/MAP ratio (0.60 ± 0.02, P < .05), drop rate, and smooth muscle/collagen ratio.Clinical TranslationThe present study provides scientific evidence for the effect of late ART on erectile function.Strengths and LimitationsThis study provides insights into the influence of late ART on erectile function through improvements in the structure of corpus cavernosum. The major limitation of this study is the difference in the time required for healing between the humans and rats, which might have a bearing on the translational relevenace of the results.ConclusionsLate ART could improve erectile function. However, as improvement requires a considerable time period, it is necessary to persist with therapy patiently for optimal results.Kataoka T, Hotta Y, Yamamoto Y, et al. Effect of Late Androgen Replacement Therapy on Erectile Function Through Structural Changes in Castrated Rats. Sex Med 2021;XX:XX–XXX.

MO056MODIFICATION OF P16, P21 AND NRF2 EXPRESSION IN A MODEL OF UREMIC VASCULAR CALCIFICATION

Abstract Background and Aims Vascular calcification (VC) is a consequence of ageing that confers development of future cardiovascular events. Accumulation of senescent cells can lead to structural and functional abnormalities in vessel wall towards increased stiffness, reduced compliance and impaired contractile and dilatory capacity. Thus, accumulation of senescent cells in the arterial wall could also contribute to the pathophysiology of VC. To test this hypothesis, the presence of cellular markers of senescence, p16, p21 and NRF2, was assessed in aorta from rat model of VC associated with chronic kidney disease (CKD). Method Six-week-old Sprague-Dawley rats underwent 5/6th subtotal nephrectomy (SNx, n=6) or no surgery (Control, n=1). After 8 weeks of renal failure, the regular chow was supplemented with high phosphate (1.2%) and vitamin D (1 µg/day, 5 days per week) for 1 or 4 weeks to initiate vascular calcification (SNx-VC group). At the end of the protocol (Figure 1), blood pressure was measured, and thoracic aorta was taken for determination of calcification by Von Kossa staining, and protein and gene expressions of p16, p21, and NRF2 by immunostaining and qPCR, respectively. Results After 4 weeks of dietary intervention, SNx rats showed an increase in pulse pressure (88±15mmHg vs 35 mmHg for the control). Marked VC was also observed in these animals, 17% of calcified area vs &amp;lt;1 % in the control rat. Calcification was focal giving strong and no calcified areas on the same aorta. Expressions of p16, p21and NRF2 proteins were enhanced at the site of calcification in the SNx-VC 4-week (4W) group whereas it was unchanged in aortic media without calcification compared to the control rat (Figure 2). In SNx-VC 1-week (1W) rats, no change in pulse pressure or vascular calcification was observed without obvious changes for staining patterns of selected markers. Using qPCR we found p16 gene expression to be higher in the most calcified aortas (4W) (fold-change = 4.32 vs 1W) and p21 gene expression to be slightly increased (fold-change = 0.53 vs 1W). NRF2 gene expression was enhanced in 1W group compared to control, but decreased in the most calcified samples (4W). Conclusion This pilot study suggests that uraemia-induced cellular senescence accompanies VCs, as suggested by the modified expression of p16 or NRF2 genes. Our observations deserve exploration in larger studies using additional senescence markers for validation. If so, cellular senescence kinetics will be evaluated in order to test whether senolytics compounds could be a therapeutic option to arrest VC in CKD.

Does dose of early intervention matter for preschoolers with externalizing behavior problems? A pilot randomized trial comparing intensive summer programming to school consultation

The goals of this study were to (a) isolate the ideal length (i.e., 4 or 8 weeks) of the Summer Treatment Program for Kindergarteners (STP-PreK) for improving school readiness and kindergarten success outcomes of preschool children with externalizing behavior problems (EBPs) during the transition to kindergarten; and (b) compare the STP-PreK model to a more standard approach in school settings (i.e., behavioral school consultation). Forty-five preschool children (82% boys; Mage = 5.16 years; 93% Hispanic/Latino background) were randomized to one of three intervention conditions: 1) 8-week STP-PreK (8W); 2) 4-week STP-PreK (4W); or 3) school year behavioral consultation (SC). Both STP-PreK groups included an 8-week parent training component. Baseline, post-intervention, and 6-month follow-up data were collected on children's school readiness and kindergarten success outcomes including parent, teacher, and objective assessment measures. Analyses using linear mixed models indicated that children's behavioral, academic, social-emotional, and self-regulation functioning significantly improved across groups. Few significant differences were found between children receiving the 4W and 8W programs, suggesting that both programs have the potential to prepare preschool children with EBP for the transition to school. Both 4W and 8W groups experienced greater initial growth across time in most domains compared to children in the SC group. However, by the end of the kindergarten year, children in the SC group caught up to children in both 4W and 8W groups on most domains. Overall, these findings suggest that all three intervention doses are effective in improving kindergarten year functioning, with some important considerations for intervention timing in preparation for the transition to elementary school. Clinical implications for school personnel are discussed.

Evaluation of hyperbaric oxygen therapy for spinal cord injury in rats with different treatment course using diffusion tensor imaging.

Animal study. To evaluate the efficacy of hyperbaric oxygen (HBO) therapy for spinal cord injury (SCI) in rats with different treatment course using diffusion tensor imaging (DTI). Hospital in Fuzhou, China. Fifty adult Sprague-Dawley rats were grouped as: (A) sham-operated group (n = 10); (B) SCI without HBO therapy group (n = 10); (C) SCI with HBO therapy for 2 weeks (SCI+HBO2W) group (n = 10); (D) SCI with HBO therapy for 4 weeks (SCI+HBO4W) group (n = 10); (E) SCI with HBO therapy for 6 weeks (SCI+HBO6W) group (n = 10). Basso Beattie Bresnahan (BBB) scores and diffusion tensor imaging parameters including fractional anisotropy (FA), mean diffusivity (MD), radial diffusion (RD), and axial diffusion (AD) values in the injury epicenter, as well as 2 mm rostral and caudal to the injury epicenter were collected and analyzed 6 weeks post-injury. Higher BBB score and FA values were found in the SCI+HBO4W group than in the SCI and SCI+HBO2W groups (all P < 0.05), whereas no significant differences of these metrics were observed between the SCI+HBO4W and SCI+HBO6W groups. MD and RD values of the SCI+HBO4W group were significantly lower than those of the SCI group (all P < 0.01). FA values were positively correlated with BBB scores. MD and RD values were negatively correlated with BBB scores. DTI parameters, especially FA, could non-invasively and quantifiably evaluate the efficacy of HBO treatment for rats with SCI and 4 weeks may be the more appropriate treatment course.

The postnatal development of perineuronal net, Nogo receptor and the effect of fluoxetine on remodeling it in the visual cortex of rats

Objective: To investigate the postnatal development of perineuronal net (PNN), Nogo receptor (Nogo R) in visual development and the effect of fluoxetine(Flx) on remodeling it in the visual cortex of adult rats. Methods: Experimental study. (1) Wistar rats were divided into postnatal weeks (PW)1,PW3,PW5,PW7,PW9 group (8 rats in each group) according to the age of PW. The changes of PNN and Nogo R were observed in the visual cortex of each group. (2) The adult rats (10 weeks after birth) were randomized into Flx 0W, Flx 2W, Flx 4W, Flx 6W and Flx 8W group (8 rats in each group) according to Flx administrational weeks. The influence of Flx on the expression of PNN and Nogo receptor in the visual cortex was detected by immunofluorescence and western blots. (3) The adult rats were randomized into Cont (negative control), Flx, binocular form deprivation(BFD,positive control) and BFD+Flx group (8 rats in each group). Flx group accepted oral administration at the dosage of 0.2 mg/ml once per day for 4 weeks. The eyelids were binocularly sutured for 2 weeks to form the BFD group, and the combination of Flx administration and BFD was performed in the BFD+Flx group.No intervention was conducted in the control group (Cont group). Immunofluorescence was used to observe the expression pattern of PNN staining by biotinylated wisteria floribunda lectin (WFA). The expression of Nogo R in the visual cortex was detected by immunofluorescence and Western blots. The expression of PNN and Nogo R were examined in each group. And t test, analysis of variance and rank sum test were employed for inter-group comparison based on the homogeneity of variance. Bonferroni method was used for multiple comparison and simple linear regression analysis was used for the trend. Results: (1) The expression of PNN (standardized b=0.97, P=0.005) and Nogo R (standardized b=0.96, P=0.010) increased during the postnatal development and the Nogo R reached the matured level at PW7 (PW7 vs. PW9, 131.83±3.78 vs. 135.11±3.92, Z=1.93, P=0.062). (2) Flx significantly decreased PNN in the visual cortex of adult rats. The density of PNN-positive cells in the visual cortex of healthy adult rats fed with Flx for 4 weeks was (86.22±7.68)/mm(2), which was similar to that of 3 weeks old rats [(84.21±6.68)/mm(2), t=2.08, P=0.073]. The expression of PNN (standardized b=-0.88, P=0.040) and it's receptor Nogo R (standardized b=-0.90, P=0.007) decreased with prolongation of Flx use. (3) The expression of Nogo R (t=13.42,11.47, 18.13; P=0.012, 0.013, 0.001; Flx, BFD and BFD+Flx group vs. Cont group) and the density of PNN (t=10.09, 7.64, 13.01; P=0.007, 0.011, 0.001; Flx, BFD and BFD+Flx group vs. Cont group) could be modulated by Flx and BFD after the critical period. There are no differences in BFD and Flx group on Nogo R changes (t=2.41, P=0.153). The expression of Nogo R protein was also different among the 4 groups (H=5.69, P=0.041). The effect of Flx combined with BFD was better than the Flx or BFD alone (Flx vs. BFD+Flx, Z=4.22, P=0.005; BFD vs. BFD+Flx, Z=3.09, P=0.010). Conclusions: The expression of PNN and Nogo R increase during the postnatal development. Chronic Flx treatment decrease the expression of PNN and Nogo R after the critical period in the visual cortex of adult rats, that is the same as BFD. (Chin J Ophthalmol, 2019, 55:37-45).

Changes and Significance of SYP and GAP-43 Expression in the Hippocampus of CIH Rats.

Synaptophysin (SYP) and growth-associated binding protein 43 (GAP-43) have been shown to be closely related to hippocampal synaptic plasticity in recent years. They are important molecular markers associated with synaptic plasticity. However, the role of SYP and GAP-43 in chronic intermittent hypoxic injury of the central nervous system needs to be further clarified. In this study, 25 adult male sprague dawley (SD) rats were randomly divided into a normal control group (CON) and a chronic intermittent hypoxia group (CIH) with four time points as follows: 1 W, 2 W, 3 W, and 4 W. The behavioural changes (primarily learning and memory abilities) were observed by the Morris water maze in each group, consisting of 5 rats per group.The localization of SYP and GAP-43 in hippocampal CA1 neurons was observed, and the expression of SYP and GAP-43 in the hippocampus was detected by Western blotting. The results showed that the mean oxygen saturation of the tail artery in CIH rats was less than that in normal rats (P < 0.05). The escape latency of CIH rats was longer than that of normal rats, and the number of space exploration platform crossings was less than that of normal rats. SYP-positive stained cells were yellow or brown and were mainly expressed on the cell membrane, while the GAP-43-positive staining was brown and was mainly expressed on the cell membrane and in the cytoplasm. The expression of SYP in plasma decreased gradually at the four time points for the CIH group (P < 0.05), while the expression of GAP-43 in the CIH 1W group increased (P < 0.05) and decreased gradually in the CIH 2 W, CIH 3 W and CIH 4 W groups (P < 0.05).

Expression of c-Jun N-terminal kinase and B cell lymphoma/leukemia-2 associated X protein in rabbit basilar artery aneurysm

Objective To observe the expression of c-Jun N-terminal kinase(JNK) and B cell lymphoma/leukemia-2 associated X protein (bax) during the formation of basilar aneurysm in the rabbit. Methods Forty New Zealand white rabbits were divided into five group, Normal group (n=5), Sham group (n=5) , Operation group(2 weeks, 8 weeks, 16 weeks)(n=10). The bilateral common carotid artery was exposed in the sham operation group by bilateral common carotid artery ligation. MRA (Magnetic Resonance Angiography) was performed at the prescribed time, All experimental animals were killed in order to observe the pathological staining of the basilar artery tip morphology, vascular intima, the middle elastic layer and the immunohistochemical staining. Results The results of immunohistochemical staining showed that the positive rate of JNK in each group was (8.600±2.074)% in the control group, (18.900±4.458)% in the 2W group (t=13.405, P=0.000), and the postoperative 8W group (25.800±3.615)% (t=22.570, P=0.000), postoperative 16W group (38.400±8.343)% (t=14.555, P=0.000); the difference between the experimental group and the control group was statistically significant. The positive expression rates of bax were: (9.400±3.050)% in postoperative 2W group, (22.100±4.748)% in postoperative 2W group (t=14.719, P=0.000) postoperative 8W group (29.200±4.894)% (t=18.866, P=0.000), postoperative 16W group (36.700±4.762)% (t=24.371, P=0.000); each experimental group was statistically significant difference from the control group. Imaging results: aneurysm The volume was: control group (7.600±3.847), (24.746±27.813) mm3,t=2.814, P=0.020), postoperative 8W group (37.083±35.695) mm3 (t=3.285, P=0.009), postoperative 16W group (105.751±97.094) mm3 (t=3.444, P=0.007); the difference between the experimental group and the control group was statistically significant. Conclusion C-Jun N-terminal kinase and bax are involved in the formation of basilar artery aneurysm in rabbits. Key words: Intracranial aneurysm; c-Jun N-terminal kinase; B cell lymphoma/leukemia-2 associated X protein

Effects of dry period length on metabolic status, fertility, udder health, and colostrum production in 2 cow breeds

Shortening the dry period improves postpartum energy balance, which has potential positive effects on metabolic health. This concept has been mainly studied in Holstein cows. The aim of this study was to assess the effect of a short dry period of 4 wk, compared with a standard dry period of 8 wk, on the metabolic status, progesterone profiles, health, and colostrum quality of dairy cows of 2 breeds, Swedish Red (SR) and Swedish Holstein (SH), not treated with antibiotics at dry off. The IgG uptake in calves was also studied to reflect the colostrum quality when shortening the dry period. Cows of both SH and SR were blocked by breed and parity and then randomly allocated to a short dry period of 4 wk (4W, n = 43) or a conventional dry period of 8 wk (8W, n = 34). Blood samples were collected wk -8, -4, -2, -1 and 1, 2, 3, 4, 6, 9, and 12 relative to calving. Prepartum, cows with a 4-wk dry period had higher concentrations of nonesterified fatty acids and lower concentrations of insulin-like growth factor-1 and insulin than 8W cows. Postpartum, plasma concentration of nonesterified fatty acids was lower, whereas plasma insulin and insulin-like growth factor-1 tended to be higher for 4W cows than for 8W cows. Plasma concentration of β-hydroxybutyrate did not differ between dry period lengths. Swedish Holstein cows with a 4W dry period responded with a lower concentration of insulin prepartum than SR and SH on an 8W dry period. The dry period length had no effect on the proportion of disturbed progesterone profiles; disturbed progesterone profiles occurred in 30% of the 4W cows and 47% of the 8W cows. In this trial, only 48.8% of the SR cows had a normal progesterone profile, which differed from the SH where 76.5% had a normal profile. Fertility-related diseases (endometritis, pyometra, anestrus, ovarian cyst) did not differ between the 2 dry period groups: 21% in the 8W group versus 12% in the 4W group, whereas mastitis tended to be more common: 26% of the 4W cows versus 9% of the 8W cows. A short dry period resulted in less colostrum but with a higher content of protein and somatic cell count. Calves were fed colostrum from their dam, and the IgG and total protein in plasma did not differ between calves to mothers with different a dry period length. Shortening the dry period could improve metabolic status in cows of both SH and SR breed postpartum, without compromising the colostrum quality. Health and progesterone profiles were not affected by the dry period length for SH or SR in this study.

Changes of two-pore K+ channel TASK-1 in diabetic myocardial injury in rats

To investigate the changes of the two- pore K+ channel TASK-1 in diabetic rats with myocardial injury. Thirty-six SD rats were divided into normal group (N), diabetes at 4 weeks (DM 4W) group, and diabetes at 8 weeks (DM 8W) group. The cardiac functions of the rats were determined using cardiac ultrasonography, and the body weight and heart weight of the rats at different time points were measured to calculate the heart/body weight ratio (HW/BW). Myocardial fibrosis in the rats was assessed using Masson's staining. The protein expression of TASK-1 in the myocardium was detected using Western blotting. Whole- cell patch clamp technique was used to record the action potential duration (APD) and twopore domain potassium channel TASK- 1 current in acutely isolated rat ventricular myocytes. meanwhile, The inhibition of TASK-1 current was observed by the TASK-1 specific inhibitor ML-365. Compared with the normal group, the diabetic rats showed significantly increased HW/BW (P &lt; 0.05), end- diastole left ventricular diameter (LVIDd), end- systolic left ventricular diameter (LVIDs), and TASK-1 protein expression, with obviously decreased left ventricular diameter shortening rate (FS) and ejection fraction (EF) (P &lt; 0.01). Masson staining showed that in diabetic rats, the collagen fibers were thickened, interwoven into a network with uneven arrangement and increased deposition. Compared with DM 4W group, the rats in DM 8W group exhibited progressive increases in LVIDd, LVIDs, HW/BW, and TASK-1 expression (P &lt; 0.01 or 0.05); FS and EF were further decreased (P &lt; 0.01). Masson staining showed worsened morphological changes of the myocardium with increased deposition. Compared with that in the normal group, the current of TASK- 1 in diabetic rats at 8 weeks was significantly reduced (P &lt; 0.01) and the duration of action potential was extended (P &lt; 0.05). The TASK-1 current was successfully inhibited by ML-365. Diabetes can induce myocardial fibrosis and aggravate myocardial injury possibly in relation to changes in the protein expression and current of the two-port potassium channel TASK-1.

Bromocriptine treatment in patients with peripartum cardiomyopathy and right ventricular dysfunction

BackgroundRight ventricular (RV) dysfunction predicts adverse outcome in peripartum cardiomyopathy (PPCM). We recently demonstrated beneficial effects associated with the prolactin release inhibitor bromocriptine at different doses when added to standard heart failure therapy in PPCM. Here, we evaluated for the first time the therapeutic potential of bromocriptine particularly in PPCM patients with RV involvement.MethodsIn this study, 40 patients with PPCM were included, of whom 24 patients had reduced RV ejection fraction (RVEF < 45%). We examined the effect of short-term (1W: bromocriptine, 2.5 mg, 7 days, n = 10) compared with long-term bromocriptine treatment (8W: 5 mg for 2 weeks followed by 2.5 mg for another 6 weeks, n = 14) in addition to guideline-based heart failure therapy in patients with an initial RVEF < 45% on the following outcomes: (1) change from baseline (Δ delta) in RVEF, (2) change from baseline in left ventricular EF (LVEF), and (3) rate of patients with full LV recovery (LVEF ≥ 50%) and (4) rate of patients with full RV recovery (RVEF ≥ 55%) at 6-month follow-up as assessed by cardiac magnetic resonance imaging.ResultsReduced RVEF at initial presentation was associated with a lower rate of full cardiac recovery at 6-month follow-up (patients with RV dysfunction: 58% vs. patients with normal RV function: 81%; p = 0.027). RVEF increased from 38 ± 7 to 53 ± 11% with a delta-RVEF of + 15 ± 12% in the 1W group, and from 35 ± 9 to 58 ± 7% with a Δ RVEF of + 23 ± 10% in the 8W group (Δ RVEF 1W vs 8W: p = 0.118). LVEF increased from 25 ± 8 to 46 ± 12% with a Δ LVEF of + 21 ± 11% in the 1W group, and from 22 ± 6 to 49 ± 10% with a Δ LVEF of + 27 ± 9% in the 8W group (Δ LVEF 1W vs 8W: p = 0.211). Full LV recovery was present in 50% of the 1W group and in 64% of the 8W group (p = 0.678). Full RV recovery was observed in 40% of the 1W group and in 79% of the 8W group (p = 0.092).ConclusionsDespite overall worse outcome in patients with RV dysfunction at baseline, bromocriptine treatment in PPCM patients with RV involvement was associated with a high rate of full RV and LV recovery, although no significant differences were observed between the short-term and long-term bromocriptine treatment regime. These findings suggest that bromocriptine in addition to standard heart failure therapy may be also effective in PPCM patients with biventricular impairment.