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Objective Nicotinamide phosphoribosyltransferase (Nampt) is a new therapeutic target for ischemic stroke. The aim of this study was to investigate protective effect of liver-derived Nampt on ischemic stroke. Methods Liver-specific Nampt knockout mice were generated using the Cre/loxP system. NamptloxP/loxP mice were crossed with liver-specific Cre recombinase expression mice (Alb-Cre), and the progeny genotypes were identified by polymerase chain reaction. Body weight of knockout mice and control mice were measured. Nampt in liver and brain was determined by Western blot assay. Middle cerebral artery occlusion (MCAO), a classical ischemic stroke model, was generated in liver-specific Nampt knockout mice and control mice by electrocoagulation. After 24 h of modeling, neurological deficit scores of each group were evaluated and TTC staining was performed to determine the cerebral infarction volume. The level of plasma Nampt in each group was determined by ELISA. Results Liver-specific Nampt knockout mice with the genotype of NamptloxP/loxPAlb-Cre were successfully constructed. The hepatic Nampt expression in knockout mice was significantly decreased by 74.2% compared to control mice, while there was no significant difference in the expression of brain Nampt protein between the knockout group and the control group. Specific knockout of liver Nampt gene expression had no effect on the body weight of mice. Under normal physiological conditions, there was no significant difference in plasma Nampt levels between liver-specific Nampt knockout mice and control mice of the same gender. 24 h after MCAO modeling, there were no significant differences in neurological deficit scores, cerebral infarct volume and plasma Nampt concentration between liver-specific Nampt knockout group and control group. Conclusion Liver-specific Nampt knockout mice are successfully constructed. Liver-derived Nampt has no significant protective effects on ischemic stroke.
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Objective To explore the expression of CRELD2 at the gene and protein levels of mouse tissues, and to provide a reference for studying the biological function of CRELD2 in various tissues. Methods The expression level of CRELD2 in the liver, pancreas, stomach, and lung of C57BL/6J mice was determined by real-time PCR and Western Blot. Results RT-PCR and WB showed that CRELD2 was expressed in mouse liver, pancreas, stomach, and lung. The relative expression levels of CRELD2 from high to low were pancreas, stomach, liver, and lung at the gene level, and pancreas, liver, stomach, and lung at protein level respectively. The result suggested that the relative expression levels of the CRELD2 gene and protein in different tissues were not completely consistent, suggesting that it is related to transcriptional regulation. Conclusion CRELD2 is expressed in mouse liver, pancreas, stomach, and lung, and the relative expression levels of CRELD2 are not completely parallel at the gene and protein level.
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Objective:To observe the optical coherence tomography angiography (OCTA) image characteristics of polypoid choroidal vascular disease (PCV) after intravitreal injection of anti-vascular endothelial growth factor drugs, and to discuss its significance in the diagnosis and follow-up of PCV.Methods:A retrospective case study. From August 2018 to January 2020, 22 eyes of 22 patients with PCV diagnosed in the ophthalmological examination of Affiliated Hospital of Weifang Medical University were included in the study. Among them, there were 10 males with 10 eyes and 12 females with 12 eyes; the average age was 67.75±9.53 years. Best corrected visual acuity (BCVA), OCTA, and indocyanine green angiography (ICGA) were performed. All the affected eyes were injected vitreously with 10 mg/ml Conbercept 0.05 ml (including Conbercept 0.5 mg) once a month for 3 consecutive months.Tthe macular area of 3 mm×3 mm and 6 mm×6 mm with an OCTA instrument was scanned, and the foveal retinal thickness (CRT) was measured, the area of abnormal branch blood vessels (BVN). pigment epithelial detachment before and 12 months after treatment (PED) height, foveal choroid thickness (SFCT) were performed. The diagnosis rate of PCV by OCTA was observed, as well as the changes of various indicators of BCVA and OCTA. Before and after treatment, BCVA and CRT were compared by paired t test; BVN area, PED height, and SFCT were compared by variance analysis. The changes in imaging characteristics of OCTA before and after treatment were analyzed. Results:Among the 22 eyes, 8 eyes were BVN; 5 eyes were polypoid lesions (polyps); 5 eyes were BVN combined with polyps; 3 eyes were not found with BVN and polyps; 1 eye with small vascular network structure, this eye was ICGA Appears as strong nodular fluorescence (polyps). The detection rate of PCV by OCTA was 86.36% (19/22). Twelve months after treatment, BVN was significantly reduced or disappeared in 16 eyes (72.72%, 16/22); polyps disappeared in 17 eyes (77.27%, 17/22). Compared with before treatment, 12 months after treatment, BCVA increased ( t=3.071), CRT decreased ( t=2.440), the difference was statistically significant ( P<0.05); the average BVN area, PED height, and SFCT decreased. The difference in average BVN area and PED height was statistically significant ( F=2.805, 3.916; P<0.05), and the difference in SFCT was not statistically significant ( F=0.047, P>0.05). Conclusions:The detection rate of PCV by OCTA is 86.36%. After PCV anti-vascular endothelial growth factor drug treatment, BVN area decrease and polyps subside. OCTA is an effective means for PCV diagnosis and follow-up after anti-VEGF drug treatment.
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Objective:To investigate the effect of endometrial thickness(EMT) on the clinical outcome of blastocyst hormone replacement freeze-thawed embryo transfer (HRT-FET) on the first progesterone day, and to analyze the threshold and optimal thickness interval corresponding to ideal clinical pregnancy rate by statistical method.Methods:The endometrial preparation protocols of 2 825 blastocyst HRT-FET cycles from January 2013 to December 2016 in Henan Provincial People′s Hospital and the Second Hospital of Hebei Medical University were studied retrospectively. According to EMT on the first progesterone day, they were divided into 5 subgroups: group Q1(EMT: 3.5-7.9 mm), group Q2(EMT: 8.0-8.9 mm), group Q3(EMT: 9.0-9.5 mm), group Q4(EMT: 9.6-10.7 mm), group Q5(EMT: 10.8-21.0 mm). Univariate analysis, classification multivariate Logistic regression analysis, curve fitting and threshold effect analysis were used to investigate the effect of endometrial thickness on clinical outcome of blastocyst HRT-FET.Results:Group Q1 was set as the control group in classification multivariate Logistic regression analysis, after adjusting for confounding factors, the clinical pregnancy rate and live birth rate in other groups were higher than the control group. The clinical pregnancy rate and live birth rate in group Q3 and Q4 were significantly increased and the differences were statistically significant(all P<0.05). The cut-off value of the endometrial thickness was 9.6 mm. When endometrial thickness was less than 9.6 mm, with 1 mm increase of endometrial thickness, the clinical pregnancy rate increased by 23%( OR=1.23, 95% CI=1.11-1.36) and the live birth rate increased by 21%( OR=1.21, 95% CI=1.10-1.33). When the endometrial thickness was thicker than the threshold, the clinical pregnancy rate did not increase significantly( OR=0.92, 95% CI=0.84-1.02), and the live birth rate showed a downward trend( OR=0.88, 95% CI=0.81-0.96). Conclusions:In the blastocyst HRT-FET cycle, endometrial thickness showes a curvilinear relationship with clinical outcome. The optimal endometrial thickness range for ideal clinical outcome is 9.0-11.0 mm.
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Objective To construct and explore the in vivo and in vitro D-galactose induced cognitive impairment models and evaluate the application value of the combined models in the study of cognitive impairments.Methods The cognitive impairment mice model induced by D-gal was prepared by continuous intraperitoneal injection of D-gal saline solution for 8weeks, followed by detection of learning and memory functions with Morris water maze.The related molecular markers in the brain tissue were assayed to evaluate the effect and application value.D-gal cell model was prepared by adding D-gal in different concentrations into the cell cultural medium of neurons harvested from the hippocampus of young mice.The effect and application value were evaluated by detecting the molecular markers related to the level of cell injury.Results The Morris water maze on the D-gal model showed that the learning and memory functions of mice in the model group were significantly lower than those in the control group.Meanwhile, the levels of apoptosis and oxidative stress in the model group were significantly higher than those in the control group.In the hippocampal neuron model of D-gal, the neurons showed a dose-dependent morphologic and functional change with the increase of D-gal dose and the levels of apoptosis and oxidative stress were significantly higher than those in the negative control.Conclusion D-galactose can be successfully used to induce cognitive impairment models both in vivo and in vitro through the decrease of the learning and memory functions of mice and induction of apoptosis and oxidative stress in neurons.Combined application of the two models of D-gal can be one of effective and promising tools for the study of cognitive impairment and pharmacodynamic evaluation.
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Several aquatic species and their enviroments were examined for presence of Vibrio parahaemo-lyticus between 2007 and 2008 in the coastal areas in Zhejiang province, and some virulence-related genes such as tdh, trh, ureC and vscC2 were investigated from the isolates. V. parahaemolyticus was recovered from 70% of the samples tested (395/566). The genes tdh, trh and ureC existed in 10.1%, 20.0% and 11.1% respectively from 395 isolates. Among the 40 tdh-positive isolates, 32.5% harbored the vscC2 gene, one of the type three secretion system 2 (T3SS2) gene family. Thirty-eight of the 40 tdh-positive isolates were positive for the Kanagawa phenomenon. Out of 44 trh-and-ureC-positive isolates, only six exhibited urease phenotype. Overall, this study reveals the significant prevalence of Vibrio parahaemolyticus in seafoods and their habitats with high diversity of virulence genes. Representative V. parahaemolyticus isolates could beused for further investigation into their pathongenecity, functional genomics, and molecular evolution.