Prevention and treatment of lipoic acid-niacin on blue-light-induced retinal damage in rats / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the preventive effect and optimal drug dose of lipoic acid-niacin(N2L)against blue light-induced retinal damage in SD rats, and to explore its possible protective mechanism.METHODS: A total of 36 specific pathogen free-grade male SD rats of 150-200 g were selected and randomly divided into normal control group, blue light injury group, N2L low-dose group(1.0 mg/kg), N2L medium-dose group(2.5 mg/kg), N2L high-dose group(5.0 mg/kg), and physiological saline group, with 6 rats in each group. The normal control group was reared in a 12 h dark and light cycle, and the rest of the groups received 9 h of daily light exposure, 3 h of blue light irradiation with a wavelength of 455 nm and an intensity of 3000±50 lx, and 12 h of darkness to establish the injury model, and were exposed to light exposure for 14 d. For 14 consecutive durations, a 1 mL dose of the corresponding drug was injected intraperitoneally. The rats were reared for another 5 d with a regular 12 h light-dark cycle and were examined by electroretinography. Specimens were prepared by over anesthesia, HE staining, and the thickness of the outer nuclear layer was observed under a optical microscope; superoxide dismutases(SOD)activity was detected by CheKineTM SOD Activity Assay Kit; and the retinal Caspase-3, quinone oxidoreductase 1(NQO1), glutathione S transferase(GST), Bcl-2, and Bax protein expression in rat retina were detected by Western blot.RESULTS: The amplitude of b-wave in dark-vision ERG 3.0 and 10.0(cd·s)/m2 stimulated light, b-wave in bright-vision ERG 3.0(cd·s)/m2 stimulated light, and the amplitude of the 2nd wave peak of oscillatory potential were significantly lower in blue light injury group than that in the normal control group(all P<0.01), while the amplitude was significantly higher in the N2L medium-dose group than in the blue light injury group(all P<0.05), and was not statistically different from that of the normal control group; the thickness of the retina in the blue light injury group was decreased in the ONL compared with that of the normal control group(P<0.001), while in the N2L medium dose group, it was thicker than that of the blue light injury group(P<0.001), and there was no statistically significant difference from the normal control group; SOD activity was significantly higher in the N2L medium-dose group than in the remaining 5 groups(P<0.05); the expression of Caspase-3, Bax, and NQO1 in the blue light injury group was higher than that of the normal control group(all P<0.01), and expression of Bax and Caspase-3 was significantly lower in the N2L medium-dose group compared with the blue light injury group(all P<0.001), whereas GST, NQO1 and Bcl-2 were significantly increased(all P<0.01).CONCLUSION:A concentration of 2.5 mg/kg N2L can effectively antagonize the damaging effect of blue light on the retina of SD rats, and it is expected to be a preventive and curative drug for it.

The role of miR-15b and vascular endothelial growth factor in the pathogenesis of novel bronchopulmonary dysplasia in neonatal rats / 中国新生儿科杂志

Objective:To study the role of miRNA-15b and vascular endothelial growth factor (VEGF) in the pathogenesis of novel bronchopulmonary dysplasia (nBPD) in rats.Methods:A total of 100 newborn SD rats were randomly assigned into BPD group and control group with 50 rats in each group. The BPD group was placed in oxygen chamber with 60% oxygen concentration and the control group received atmospheric air. The morphological changes of lung tissues were observed on 1 d, 7 d, 14 d and 21 d and the radial alveolar counts (RAC) and alveolar septal thickness (AST) were measured. The expression of miR-15b was measured using real-time quantitative PCR and the expression of VEGF in lung tissue was examined using ELISA method.Results:With prolonged oxygen exposure, the lung tissue of the BPD group showed a decrease in the number of alveoli, a gradual loss of the normal structure of alveoli and a significant widening of the alveolar septum. On 7 d, 14 d and 21 d, RAC values [(6.19±0.29) vs. (6.86±0.92), (5.35±0.67) vs.(9.75±0.34), (3.96±0.45) vs. (10.04±0.52)] were significantly lower in the BPD group than the control group ( P<0.05). On 7 d, 14 d and 21 d,the levels of AST in BPD group were significantly higher than the control group [(6.87±0.41) μm vs. (6.43±0.31) μm, (8.94±0.25) μm vs. (5.36±0.26) μm, (9.61±0.30) μm vs. (4.55±0.32) μm] ( P<0.05). On 7 d, 14 d and 21 d,the miR-15b expression in BPD group were significantly higher than the control group [(1.12±0.11) vs. (0.84±0.09), (1.33±0.09) vs. (0.73±0.07), (1.66±0.15) vs. (0.45±0.10)] ( P<0.05).On 7 d, 14 d and 21 d, VEGF in BPD group were significantly lower than the control group [(10.89±1.67) pg/ml vs. (23.86±4.38) pg/ml, (8.75±1.28) pg/ml vs. (53.94±3.49) pg/ml, (4.66±1.12) pg/ml vs. (70.37±3.10) pg/ml] ( P<0.05). Conclusions:MiR-15b and VEGF may play a role in the development of nBPD.

" Multi-hits" based SD rat models of cognitive dysfunction and research on related mechanisms / 中华实用儿科临床杂志

Objective:To establish Sprague-Dawley (SD) rat models of cognitive impairment through repeated stimulation of lipopolysaccharide (LPS) in the early brain development, and to inquire into the effect of " multi-hits" mediated by inflammatory response on the histology and behavior of SD rat models and related molecular mechanisms.Methods:This study adopted a group design for experiments.The " multi-hits" SD rat models were established by intraperitoneal injection of LPS.According to the random number table method, 24 pregnant rats were randomly divided into 4 groups: control group, LPS1 group, LPS2 group and LPS3 group, 6 rats in each group.In the control group, saline was intraperitoneally injected into rats with gestational age of 18 days and 20-day-old neonatal rats.Rats with gestational age of 18 days were intraperitoneally injected with saline in the LPS1 group, 0.05 mg/kg LPS in the LPS2 group, and 0.1 mg/kg LPS in the LPS3 group.The pups in LPS1-3 groups were all injected intraperitoneally with 1 mg/kg LPS at the postnatal age of 20 days.The motor and cognitive function of the pups were evaluated overall by behavioral experiments such as forelimb suspension tests, grid tests and water maze tests.The relative expression of glial fibrillary acidic protein (GFAP), Notch1 and Jagged1 in brain tissue of pups was mainly detected by Western blot (WB) and histological experiments.One-way ANOVA analysis of variance and independent samples t- test were used to compare data among groups and between groups, respectively. Results:(1) Behavioral experiments: compared with the control group, LPS1-3 groups showed progressive decrease in forelimb suspension time [(34.81±5.66) s, (22.47±4.35) s, and (13.20±4.25) s vs.(43.88 ± 4.85) s], and the number of missteps in the grid experiment increased progressively (16.13±2.90, 20.75±3.10, 25.13±4.45 vs.9.00±2.72). The differences were statistically significant ( F=69.77, 35.59, all P<0.001). Both the escape latency and total distance in Morri′s water maze test increased progressively ( P<0.05). (2) WB experiment: the relative expression levels of GFAP, Notch1 and Jagged1 proteins in LPS1-3 groups were significantly higher than that in the control group ( P<0.05). (3) Hematoxylin-eosin (HE) staining and electron microscope pathology: compared with the control group, LPS1-3 groups had more loosely arranged frontal cortices and more obvious cell pyknosis.Under the electron microscope, the cytoplasm was swelling to varying degrees, mitochondrial cristae were broken, and part of the nuclear membrane was damaged. Conclusions:In the " multi-hits" cognitive impairment model, the damage to the brain tissue structure and behavioral changes of pups may be related to the up-regulation of Notch1/Jagged1 pathway mediated by repeated exposure to LPS.

Effect of operative trauma and multiple propofol anesthesia on neurodevelopment and cognitive function in developmental rats / 浙江大学学报·医学版

To investigate the effect of multiple propofol anesthesia and operative trauma on neuroinflammation and cognitive function in development rats and its mechanism. A total of 104 13-day-old neonatal Sprague-Dawley rats were randomly divided into 4 groups with 26 rats in each group: control group was treated with saline q.d for propofol group was treated with propofol q.d for surgery group received abdominal surgery under local anesthesia and then treated with saline q.d for surgery with propofol group received propofol anesthesia plus abdominal surgery under local anesthesia with ropivacaine at d1, then treated with propofol q.d for At d2 of experiment, 13 rats from each group were sacrificed and brain tissue samples were taken, the concentration of TNF-α in hippocampus was detected with ELISA, the expression of caspase-3 and c-fos in hippocampal tissue was determined with immunohistochemical method, the number of apoptotic neurons in hippocampus was examined with TUNEL assay. Morris water maze test was used to examine the cognitive function of the rest rats at the age of 60 d, and the TNF-α concentration, caspase-3, c-fos expressions and the number of apoptotic neurons in hippocampus were also detected. Compared with control group, TNF-α concentration, caspase-3, c-fos expression and the neuroapoptosis in hippocampus increased significantly in other three groups (all 0.05). Morris water maze test showed that there were no significant differences in swimming speed, escape latency, target quadrant residence time and crossing times among groups (all >0.05). TNF-α level, expressions of caspase-3 and c-fos and apoptotic cell numbers in hippocampus had no significant differences among the 4 adult rats groups (all >0.05). Abdominal surgery and multiple propofol treatment can induce neuroinflammation and neuroapoptosis in hippocampus of neonatal rats, however, which may not cause adverse effects on neurodevelopment and cognitive function when they grown up.

Salt-inducible kinase 2 regulates energy metabolism in rats with cerebral ischemia-reperfusion / 浙江大学学报·医学版

To investigate the effects of salt-inducible kinase 2 (SIK2) on energy metabolism in rats with cerebral ischemia-reperfusion. Adult SD male rats were divided into 5 groups: sham group, ischemia group, reperfusion group, adenovirus no-load group, and SIK2 overexpression group with 5 animals in each group. The middle cerebral artery occlusion (MCAO) was induced with the modified Zea-Longa line thrombus method to establish the cerebral ischemia reperfusion model. Eight days before the MCAO, SIK2 overexpression was induced by injecting 7 μL adenovirus in the right ventricle, then MCAO was performed for followed by reperfusion HE staining was used to observe the pathological changes of cerebral tissue in rats; TTC staining was used to observe the volume of cerebral infarct. The levels of adenosine triphosphate (ATP) and adenosine diphosphate (ADP) in rat brain tissue were detected by ELISA; the levels of SIK2 and hypoxia-inducible factor 1α (HIF-1α) in the rat brain tissues were detected by RT-qPCR and Western blotting. Compared with the sham group, SIK2 level was decreased in the ischemia group, and it was further declined in the reperfusion group (<0.05). Compared with the sham group and ischemic group, the pathological injury in reperfusion group were more severe, and the infarct size was larger; compared with the reperfusion group and adenovirus no-load group, the pathological injury of the SIK2 overexpression group was milder, and the infarct size is less. Compared with the sharn group, HIF-1α was increased in both ischemia group and reperfusion group, especially in ischemia group (all <0.05); HIF-1α level in the SIK2 overexpression group was higher than that in the reperfusion group and adenovirus no-load group (all <0.05). ATP level in ischemia group and reperfusion group was lower than that in the sham group, and the reperfusion group decreased more significantly than the ischemia group (<0.05); ADP content was increased in the ischemia and reperfusion group, and the ADP content in reperfusion group was significantly higher than that in the ischemia group (<0.05). ATP level in the SIK2 overexpression group was higher than that in the reperfusion group and adenovirus no-load group (all <0.05), and ADP was decreased in the SIK2 overexpression group (all <0.05). SIK2 can up-regulate the ATP level and down-regulate the ADP level in rat brain tissue and alleviate cerebral ischemia-reperfusion injury by increase the level of HIF-1α.

Effect of electro-acupuncture therapy on limb spasm and excitability of motor neurons in stroke rats / 浙江大学学报·医学版

To investigate the effect of electro-acupuncture therapy on limb spasm and excitability of motor neurons in stroke rats. Ischemic stroke model was induced with middle cerebral artery embolization in SD rats. Thirty-three modeled rats were randomly divided into model group, electro-acupuncture group, and baclofen group with 11 rats in each group, and another 10 rats were taken as sham operation group. The electro-acupuncture group and the baclofen group were treated with electro-acupuncture and baclofen tablets respectively. The model group and the sham operation group had no intervention. The neural function was evaluated with Bederson's scale and balance beam test; the muscle tension was measured with electrophysiography; the pathological changes of brain tissue was examined with HE staining; the content of glutamic acid (Glu) and γ-aminobutyric acid (GABA) in rat cerebral cortex was analyze with enzyme linked immunosorbent assay (ELISA) method, the expression of metabotropic glutamate receptor 1a () and γ-aminobutyric acid type B receptor subunit 1 () mRNA were detected with RT-qPCR. Compared with the model group, the neurological function scores of the electro-acupuncture group and the baclofen group showed a downward trend at d7 after operation (all >0.05), and the neurological function scores of the electro-acupuncture group and the baclofen group were significantly decreased at d12 after the operation (all 0.05). Compared with the model group, the electrophysiological results of the electro-acupuncture group and baclofen group were significantly increased after operation (all <0.05). The results of HE staining showed that there was no cell edema and degeneration in the sham operation group, no pyknosis of the nucleus, and no bleeding in the interstitium. Cell edema and degeneration and mesenchymal congestion appeared in the model group. Compared with the model group, the cytoplasmic edema and degeneration and the interstitial bleeding in the electroacupuncture group and the baclofen group were reduced. Compared with sham operation group, the Glu content and the relative expression of mRNA was increased in the model group, electro-acupuncture group and baclofen group, while the GABA content and the relative expression of mRNA decreased (all <0.05). Compared with model group, the Glu content and the relative expression of mRNA in the electro-acupuncture group and baclofen group decreased, and the GABA content and relative expression of mRNA increased (all <0.05). Electro-acupuncture may improve limb spasm after stroke through regulating the expression of Glu and GABA in the cerebral cortex and the excitability of motor neurons in rats.

Disorder of intestinal amino acid pathway in depression-like offspring rats induced by maternal separation / 浙江大学学报·医学版

To investigate the intestinal amino acids pathway in depression-like offspring rats induced by maternal separation. Sprague-Dawley (SD) female rats were randomly divided into a control group (=8) and a maternal separation group (=8). After normal delivery, the maternal rats were separated from offsprings for 14 consecutive days and 3 h per day in maternal separation group; while rats in the control group was received no interventions in postpartum. Depression-like behaviors of offspring rats were evaluated using the sucrose preference test, novelty suppressed feeding test, and forced swimming test. Amino acid analyzer was used to detect the changes of amino acid contents in the small intestine, and the expressions of alanine-serine-cysteine transporter 2 (ASCT2), solute carrier superfamily 6 member 19 (BAT1) and L-type amino acid transporter 1(LAT1) were detected by Western blot. The weight of the offspring rats in the maternal separation group was significantly lower than that of the control group at 21 and 28 d (=4.925 and 5.766, all <0.01). Compared with the control group, the percentage of sucrose preference of the offspring rats in the maternal separation group was significantly reduced (=2.709, <0.05), and the feeding latency was significantly prolonged (=-13.431, <0.01). The immobility time in FST of maternal separation group was significantly longer (=-3.616, <0.01).Increased concentration of aspartic acid (=-6.672, <0.01) and down-regulation of glutamine (=3.107, <0.01) and glycine (=9.781, <0.01) were observed in maternal separation group. Western blot analysis revealed that the protein expressions of ASCT2 (=6.734, <0.01) and BAT1 (=9.015, <0.01) in maternal separation group were reduced, while the expression of LAT1 was increased (=-8.942, <0.01). Maternal separation can induce the depression-like behavior in offspring rats; the amino acid contents and the amino acid transporter expression in the small intestine are reduced, which may be related to depression-like behavior induced by maternal separation.

Changes of pdgfr-β+ microvascular wall cells in blood-brain barrier permeability of SD rats after status epilepticus / 国际儿科学杂志

Objective To investigate the blood-brain barrier permeability of platelet-derived growth factor receptor-β + (pdgfi-β +) vascular wall cells at 2h,24h,3 d,7 d,14 d and 21 d after lithium-pilocarpine-inducedstatus epilepticus.Methods One hundred and thirty-five clean male Sprague-Dawley rats were randomly divided into control group (n =15) and model group (n =120).The model group was divided into 2h,24h,3d,7d,14d and21d after SE.We evaluated the permeability of blood-brain barrier in hippocampus of rats in each group after status epilepticus by Evans blue method and wet and dry weight method.We observed the ultrastructural changes of pericytes and blood-brain barriers at different stages after onset by electron microscopy.We used Western Blot to detect the expression of pericyte marker pdgfr-β and α-SMA in hippocampus at different stages after onset.Results (1) The blood-brain barrier permeability increased after epileptic seizures (P ＜ 0.05),and the permeability was the highest at 24h after onset (P ＜ 0.01),and gradually returned to normal after 3d and later.(2) Transmission electron microscopy showed that the ultrastructure of cerebral microvascular pericytes and their basement membranes were degenerated after SE.(3) Western blot showed that the expression level of pdgfr-β and α-SMA at 24 h after SE was significantly higher than that of the control group (P ＜0.05),and gradually became stable after 3d and later.Conclusion Pdgfr-β + microvascular wall cells in brain microvessels may be involved in the opening of blood-brain barrier after status epilepticus,and may be dedicated to the conversion of disease into refractory epilepsy.

The change of COX-2 expression in rats with stress urinary incontinence and its significance / 医学研究生学报

Objective　There are a few researches on the mechanism of stress urinary incontinence (SUI). The article aimed to examine the changes of COX-2 expression in the urethra, vagina and urethral smooth muscle of SUI rat mode to evaluate the effect of estrogen on COX-2 expression. Methods　Sixty unbearing healthy female SD rats and fifteen male SD rats were gathered for spontaneous delivery. SUI rat models were constructed using expanded vagina, expanded vagina + ovariectomy respectively after delivery, which were expanded vagina group and expanded vagina + ovariectomy group. Six successfully modeled rats were chosen for the follow-up experiment. SD rats modeled after normal pregnancy were the control group. Sneezing experiment and urodynamic examination were used to examine the maximum bladder capacity (MBC) and abdominal leak point pressure (ALPP). Fluorescent quantitative PCR and western blot were applied to detect the expressions of COX-2 mRNA and protein, and immunohistochemistry was used to detect the expressions of COX-2 in urethra, vagina and urethral smooth muscle. Results Compared with control group, ALPP in two experimental groups were significantly decreased, among which ALPP in expanded vagina + ovariectomy group was significantly decreased in comparison to expanded vagina group(P<0.05). Compared with control group, the expressions of COX-2 mRNA and protein in expanded vagina group and expanded vagina+ovariectomy group were significantly higher, among which the figures in expanded vagina+ovariectomy group were significantly higher than those in expanded vagina group(P<0.05). The result of immunohistochemistry showed staining intensity integral expression of COX-2 in vaginal tissues of control group, expanded vagina group and expanded vagina+ovariectomy group were 0.50±0.54, 5.55±0.54, 9.33±0.81, so differences between any two groups were of statistical significance(P<0.05); staining intensity integral expression of COX-2 in urethral smooth muscle of control group, expanded vagina group and expanded vagina+ovariectomy group were 0.66±0.51, 5.33±0.51, 8.50±0.54, so differences between any two groups were of statistical significance (P<0.05). Conclusion The expression of COX-2 was related to the mechanism of SUI. The decrease of estrogen may increase the expression of COX-2 in SUI rats, which supports the treatment of SUI.

Chronic Toxicity of Agarwood Extracts Produced by Agar-Wit on SD Rats / 中国药学杂志

OBJECTIVE: To assess the chronic toxicity of agarwood extracts produced by agar-wit. METHODS: Agarwood usage in Chinese Pharmacopoeia was referred. SD rats were used as the experimental animal and fed with agarwood extracts at pharmaceutical dosages (1 262.4, 631.2 and 157.8 mg•kg-1) and functional food dosages (150, 105 and 45 mg•kg-1) for 90 d. The weight and food utilization were recorded. The rats were killed on the 90th day, blood and urine were collected for routine blood, serum biochemical indicators and urine index tests, and the bodies were dissected for histological examination. RESULTS: The two groups of rats showed same results. The body weight, food utilization, routine blood, serum biochemical indicators and urine index were not significantly changed compared with the control group (P>0.05). Histological examination showed that the male and female rats both had some lesions, but the group of high dose extracts had no significant difference compared with the control group, also indicating that the lesions had no relationship with agarwood extracts. CONCLUSION: The agarwood extracts produced by agar-wit have no chronic toxicity for mammal in regards of body weight, biochemical indicators and pathology at both pharmaceutical dosages and functional food dosages.

Structural and functional changes of immune system in natural aging SD rats / 中国实验动物学报

Objective To investigate the structural and functional changes of immune system in aging Sprague-Dawley rats. Methods Sixty SPF 4-6-week old SD rats(male:female=1:1)were used in this study. Ten of them were randomly taken and euthanized every 6 months. The dynamic changes of T cell proliferation,expression of cytokines,serum levels of SOD and MDA and histopathology were examined. Results Obvious histological changes of thymus and spleen were observed in the aging rats. Compared with the young SD rats,in the aging rats,the lymphocyte transformation ability was decreased(P<0.05,P<0.01),number of splenic cells was declined(P<0.05,P<0.01),NK(P<0.05,P<0.01)and T cell subset was reduced(P<0.05,P<0.01),and the production of IL-2 was decreased as well(P<0.05, P<0.01). The serum level of SOD in old rats was lower,MDA was increased,with significant differences(P<0.05,P<0.01). The immunohistochemical staining showed that more extensive staining was found in the nuclei of thymocytes from the aging rats,while the 8-OHdG formation in thymic tissues was mostly located in the thymic medulla. Conclusions Aging process is accompanied by immune impairment,oxidative stress can also impair the immune response in aging rats. Our findings indicate that structural and functional alterations of immune system in aging rats may be closely related with ox-idative damages.

Swimming plus medication reduces the expressions of cytokines in rats with chronic abacterial prostatitis / 中华男科学杂志

Objective@#To observe the effects of swimming plus medication on the expressions of cytokines in rats with chronic abacterial prostatitis (CAP).@*METHODS@#Forty healthy adult male SD rats were randomly divided into five groups of equal number, normal control, CAP model control, medication, exercise therapy, and exercise + medication. The CAP model was made by Xiaozhiling injection, and at 7 days after modeling, the rats in the medication and exercise + medication groups were treated intragastrically with Qianlie Shutong Capsules (0.016 g/ml) at 20 ml per kg of the body weight qd, those in the exercise therapy and exercise + medication groups were made swim at a regular time once a day, 35 minutes on the first day and 5 minutes more on the second until 50 minutes once, for 4 successive weeks, and those in the normal control, model control and exercise therapy groups received normal saline only. After 14 and 28 days of treatment, all the rats were killed and their prostates harvested for observation of histopathological changes and determination of the expressions of TNF- α, IL-1β and IL-6 in the prostatic tissue homogenate by ELISA.@*RESULTS@#After 14 days of treatment, the expression levels of TNF-α, IL-1β and IL-6 were significantly elevated in the groups of CAP model control (［183.08±8.07］ pg/ml, ［57.55±3.53］ pg/ml and ［256.15±13.95］ ng/L), medication (［118.49±8.06］ pg/ml, ［42.64±4.64 ］ pg/ml and ［200.74±9.33］ ng/L), exercise therapy (［169.63±10.64］ pg/ml, ［50.45±5.71］ pg/ml and ［245.23±6.49］ ng/L), and exercise + medication (［107.82±7.81］ pg/ml, ［40.35±6.93］ pg/ml and ［187.04±10.85］ ng/L) as compared with those in the normal control (［20.36±1.82］ pg/ml, ［14.64±1.91］ pg/ml and ［70.58±2.09］ ng/L) (P<0.05). At 28 days, the levels of TNF- α, IL-1β, IL-6 were remarkably lower in the exercise + medication group (［29.30±3.78］ pg/ml, ［16.91±1.24］ pg/ml and ［ 88.65±6.74］ ng/L) than in the medication group (［39.67±3.19］ pg/ml, ［26.27±3.49］ pg/ml and ［110.26±6.33］ ng/L) (P<0.05) and close to those of the normal control group (［19.34±1.76］ pg/ml, ［13.68±1.06］ pg/ml and ［71.34±2.50］ ng/L). During the treatment, no obvious pathological changes were found in the prostate tissue of the normal control rats, while significant chronic prostatic inflammation was observed in the CAP models, and the inflammation was relieved in different degrees after intervention, most significantly in the exercise + medication group.@*CONCLUSIONS@#Swimming can relieve prostatic inflammation and swimming plus medication can effectively reduce the expressions of cytokines and alleviate histological damage in the prostatic tissue of CAP rats.

Protective effect of astaxanthin against epididymal oxidative damagein rats with ornidazole-induced oligoasthenozoospermia / 中华男科学杂志

Objective@#To investigate the improving effect of astaxanthin (AST) on the sperm quality of rats with ornidazole (ORN)-induced oligoasthenozoospermiaand its action mechanism.@*METHODS@#Forty adult male SD rats were equally randomized into groups A (solvent control), B (low-dose ORN ［400 mg/（kg·d）］), C (high-dose ORN ［800 mg/（kg·d）］), D (low-dose ORN ［400 mg/（kg·d）］ + AST ［20 mg/（kg·d）］), and E (high-dose ORN ［800 mg/（kg·d）］ + AST ［20 mg/（kg·d）］), all treated intragastrically for3 weeks.After treatment, the epididymal tails ononeside was taken for determination of sperm concentration and activity, and the epididymideson the other side harvested for measurement of the activities of GSH-Px, GR, CAT and SOD and the MDA contentin the homogenate.@*RESULTS@#Compared with group A, sperm motilityin the epididymal tail andGSH-Px and SOD activities in theepididymiswere markedly decreased while the MDAcontent significantlyincreased in group B (P<0.05), spermmotility and concentrationin the epididymal tail, testisindex, and the activities of GSH-Px, GR, CAT and SOD in the epididymis were remarkably reduced while theMDA contentsignificantly increased in group C(P<0.05). In comparison with group B, group D showed markedly increased sperm motility (［45.3±8.7］% vs ［66.3±8.9］%, P<0.05) in the epididymal tail and SOD activity in the epididymis (［116.7±25.3］ U/mg prot vs ［146.1±23.8］ U/mg prot, P<0.05), decreased MDA content(［1.68±0.45］ nmol/mg prot vs ［1.19±0.42］ nmol/mg prot, P<0.05).Compared with group C, group Eexhibited significant increases in the weight gained (［89.0±9.5］ vs ［99.9±4.1］ %, P<0.05) and sperm motility (［17.9±3.5］% vs ［27.3±5.3］ %, P<0.05) but a decrease in the content of MDA (［2.03±0.30］ nmol/mg prot vs ［1.52±0.41］ nmol/mg prot, P<0.05).@*CONCLUSIONS@#AST can improve spermquality in rats with ORN-inducedoligoasthenozoospermia, which may be associated with its enhancing effect on the antioxidant capacity of the epididymis.

A comparative study about the time of pregnancy and gestational age of SD rats by three methods / 中国基层医药

Objective To explore more accurate method about the time of pregnancy and gestational age of SD rats.Methods 7 to 8-week-old SD rats (18 females,6 males) were selected,the ratios of male to female were 2:1,5:2,1:1 and 4:1.Through vaginal plug observation,vaginal smear method,weight comparison method to verify the exact date of pregnancy in the next morning.Results Vaginal observed at milky vaginal plug sperm could be observed as sickle rat sperm head.Body weight of pregnant rats changed obviously.Three detection methods were used to detect the pregnancy time for 18 females.The positive rate of vaginal plug observation was 5.6％.The positive rate of vaginal smear method was 44.4％.The false negative rate of vaginal smear method was 11.1％.The positive rate of body weight method was 11.1％.The false positive rate was 5.6％.Conclusion The accuracy of vaginal smear is higher than the vaginal plug method and weight comparison method,the vaginal plug method can be used as the premise of the other two methods.Comparison method can be used as supplementary that the other two methods missed.

Distribution of autofluorescent microorganisms in the rat intestine / 中国实验动物学报

Objective To study the distribution of intestinal autofluorescent microorganisms in the rat intestine at different developmental stages. Methods The distribution of intestinal autofluorescent microorganisms in rat intestine at va-rious developmental stages was tested and evaluated using a small animals living imaging system. First, standard E. coli strain was tested by fluorescence detection in vitro. Then, the distribution of E. coli under the same test conditions was tested. The intestinal autofluorescent bacteria distribution was detected in the SD rats at 3 days,14 days and 60 days of age. After expanding the range of excitation wavelength fluorescence detection,removing the background of fluorescence feed and feces and other foreign autofluorescent substances. Results E. coli can be excited in the range of 485 -535 nm wave?length and to emit fluorescence. E. coli mainly existed in the stomach and only a few E. coli were found in the ileum of 3?days old SD rat. . In the 14?days old rats, E. coli mainly existed in the stomach and cecum, and only a few E. coli were found in the ileum. In the 60-days old SD rats, E. coli mainly existed in the ileum, and only a few E. coli were found in the colon, cecum and jejunum. After the expansion of the excitation light wavelength range of fluorescence detection, E. co?li were observed mainly in the ileum, and only a few E. coli were found in the stomach in 3?days old SD rat. E. coli mainly existed in the stomach, then the cecum and only a few E. coli were found in the ileum and jejunum in 14-days old SD rats. E. coli could be found in the whole intestinal system but mainly in the ileum and cecumin of the 60-days old rats. Conclu?sions Examining the intestinal autofluorescent microbes with the small animal in vivo imaging system can be helpful and make guidance to study the distribution of intestinal microbes in the host at different developmental stages, and to provide a basis for studying the relationship of intestinal microbes with its host and the gastrointestinal drug administration.

Prevention of the development of third molars in SD rats / 实用口腔医学杂志

Objective:To observe the effect of dexamethasone(DXM) in the prevention of the development of third molars in SD rats.Methods:12 SD rats of 5 days old(PN 5 d) and 12 SD rats of 21 days old(PN 21 d) were respectively divided into 3 groups(n =4).The rats were injected with 0.1 ml DXM at 1.94 × 10-3 mmol/L,1.94 × 10-4 mmol/L and 1.94 × 10-5 mmol/L respectively into the experimental side,and 0.1 ml normal saline into the control side.PN5d SD rats were respectively killed 21 d and 43 d after injection.PN 21 d SD rats were killed 43 and 52 d after infecion.The PN 21 d SD rats injected with the highest concentration of DXM were the successful models of development prevemion of their third molars in experimental side.The tissue samples were subjected to general observation,radiographic and HE staining examination.Results:After injection,in the highest concentration groups,third molars on the experimental sides of PN 21 d SD rats were not erupted,the control side showed normal eruption.X-ray examination showed crown formation on the experimental sides of the third molars,but without root formation.HE biopsy revealed disorder of root development.Conclustion:DXM with certain concentration injected into local area in late bell stage can prevent tooth development.

Establishment of male rat models for fertility after liver transplantation / 器官移植

Objective To establish male rat models for fertility following liver transplantation. Methods Male Sprague-Dawley (SD) rats were used as the donors and recipients of liver transplantation. The donor liver was transplanted with two-cuff technique. Liver transplantation was performed in 15 male SD rats. At 3 weeks after liver transplantation, 5 rats were randomly sacrificed for detection of sperm deformity rate. The remaining male rats were mixed bred and mated with healthy female SD rats at a ratio of 1︰2. General conditions of the rats undergoing liver transplantation were recorded. Liver function parameters were detected after liver transplantation. Postoperative sperm deformity rate was observed. The pregnant status of female rats and health situation of their offsprings was monitored. Results All 15 rats (100%) underwent liver transplantation successfully. Nine rats (9/10) survived longer than 8 weeks. Liver function parameters were normal in male rats following liver transplantation. The sperm deformity rate was ranged from 0.5% to 1.3%. Ten male rats undergoing liver transplantation were mixed bred with female rats at a ratio of 1︰2 for 1 week. All female rats were successfully mated and delivered their offsprings after 3 weeks. The offsprings had no evident physiological deformity. Conclusions Male rat models for fertility are successfully established after liver transplantation, which serve as an animal model to evaluate the fertility performance in male patients undergoing liver transplantation.

An experimental study of isolation of SD rat adipose-derived stem cells using suspended explant culture method / 中国免疫学杂志

Objective:To isolate SD rat adipose-derived stem cells(ASCs)by suspended explant culture method.Methods: The healthy rat inguinal fat pads were obtained.The SD rat ASCs were isolated by suspended explant culture method and explant adherent culture method.The growth status and morphology were observed.The growth curve and cell surface markers CD29,CD44,CD45 of the 3rd passage cells were analyzed respectively by CCK-8,immunocytochemistry;the 3rd passage cells were induced individually by adipogenic differentiation medium and osteogenic differentiation medium,and cells were examined by oil red O staining and alizarin red staining.Results: The SD rat ASCs obtained by the two methods exhibited a spindle-shaped appearance and could rapidly expand.The cell growth curves were typical of S type.Immunocytochemistry analysis revealed that the third passage of SD rat ASCs were positive for CD29,CD44,but were negative for CD45.SD rat ASCs were positive for oil red O staining at 14 days after adipogenic induction,and positive for alizarin red staining at 14 days after osteogenic induction.Conclusion: Isolation of SD rat ASCs by suspended explant culture method is successfully established.The method is simple.It provides a new method for the isolation of SD rat ASCs in vitro.

Effects and possible mechanism of sphingosine-1-phosphate-stimulated insulin secretion from rat islets / 中国药理学通报

Aim To observe the effects of glucose-stim-ulated insulin secretion ( GSIS ) on rat islets after S1 P treatments and the underlying molecular mechanisms. Methods Collagenase P and Histopaque 1077 were used to digest and isolate rat pancreatic islets, and Dispase II was used to digest pancreatic islet to obtain pancreatic cells. Insulin secretions were measured after S1P (0~20 μmol·L-1 ) treatment under low glucose ( LG, 2. 8 mmol·L-1 ) and high glucose ( HG, 16. 7 mmol·L-1 ) conditions. Patch-clamp recordings were applied to monitor voltage-dependent potassium chan-nel currents (Kv currents) after S1P treatment. Calci-um image technique was used to measure the changes of intracellular Ca2+ concentration after S1P ( 0 ~20μmol·L-1 ) treatments. Results HG group signifi-cantly increased insulin secretion compared to LG group ( P0. 05 ) . S1 P increased insulin secretion significantly in a dose-dependent man-ner under HG condition ( P<0. 01 ) . Kv currents ofβcells were inhibited significantly after S1 P treatment ( P<0. 01 ) . S1 P increased the concentrations of in-tracellular Ca2+ in a dose-dependent manner under HG condition( P <0. 01) . Conclusion S1P may pro-mote GSIS by inhibiting Kv currents and increasing the level of intracellular Ca2+.

Comparison of the repeated dose toxicity of doxorubicin liposome injection and doxorubicin injection in rats / 中国比较医学杂志

Objective To investigate the repeated dose toxicity of doxorubicin liposome injection and doxorubicin injection in rats. Methods Ninety SD rats ( body weight 180-220 g, male:female=1:1 ) were divided into 3 groups (30 rats in each group), and were administered intravenously with physiological saline, doxorubicin liposome injection (1 mg·kg-1 ) and doxorubicin injection ( 1 mg·kg-1 ) , respectively, once every three days for thirteen times. The body weight, blood biochemistry, hematology, organ coefficient and histopathology were analyzed for the overall toxicity assessment. Results The rats administered with doxorubicin liposome injection (1 mg/kg) showed hair loss and skin ulcer, significantly reduced growth of body weight, increased levels of urea nitrogen ( BUN ) , alanine aminotransferase ( ALT ) , blood platelet ( PLT ) , and kidney and heart coefficients, decreased thymus and testicular coefficients, myofibrillar rupture and lysis, and partial loss of cell nuclei, hyaline casts in the renal convoluted tubules, interstitial edema and loss of spermatogenic cells in the testicular tubules. Compared with the doxorubicin liposome injection group, similar abnormal changes were also observed in the doxorubicin injection group, but the hair loss and skin ulcer were milder and the heart and kidney toxicities were severer. Conclusions Compared with doxorubicin injection, the doxorubicin liposome injection causes milder heart and kidney toxicity but more serious skin toxicity.