The Expression of Cyclins in Neurons of Rats after Focal Cerebral Ischemia

The change of the expression of Cyclins in neurons of rats after focal cerebral ischemia was investigated. Ischemia was induced by temporary middle cerebral artery occlusion （MCAO）. The experimental rats induced by MCAO were sacrificed on 7th and 14th day after reperfusion. The brain was taken out at 7th and 14th day after injury, and the expression of Cyclin D1, E, A and B1 in neurons of cerebral cortex or hippocampal CA1 region was detected by immunofluorescence and confocal microscope. The results showed that after MCAO, in the ipsilateral CA1 subfield of hippocampus the expression of Cyclin D1, E, A and B1 in neurons was significantly gradually up-regulated at 7th and 14th day after reperfusion （P〈0.05） as compared with that in control group. In the ipsilateral cerebral cortex the expression of Cyclin D1 and B1 in neurons was notably gradually down-regulated at 7th and 14th day, and that of Cyclin E and A was significantly up-regulated at 14th day after reperfusion as compared with that in control group （all P〈0.05）. It was concluded that there was a differential sensitivity among neurons from different brain regions to ischemic injury. But all of them re-enter into cell cycle after MCAO.     

Effect of Ovariectomy on Serum Adiponectin Levels and Visceral Fat in Rats

This study was aimed to examine the effect of ovariectomy on visceral fat, serum adi- ponectin levels and lipid profile. Forty-five female Sprague DawIey rats were divided into three groups （n=15 each）： ovariectomized group （OVX）, ovariectomized plus estrogen-treated group （OVX＋E2）, and sham-operated group （SHAM）. Body weight, abdominal adipose tissues, serum adiponectin and lipid profile were measured and compared among the groups after three-month feeding post-surgery. Signifi- cant increases in body weight and visceral fat were found in ovariectomized rats when compared with sham-operated ones and significant increases were also observed in serum adiponectin, triglyceride and very low density lipoprotein cholesterol levels in ovariectomized rats. Body weight, visceral fat and se- rum adiponectin levels were profoundly reduced in OVX＋E2 group as compared with OVX group. It was concluded that ovarian hormone deficiency induced by ovariectomy leads to significant increases in body weight and visceral fat, along with increased serum adiponectin, triglyceride and very low density lipoprotein cholesterol levels in rats. Attenuation in these changes can be achieved by estrogen supple- mentation.     

Observation of Post-MCAO Cortical Inflammatory Edema in Rats by 7.0Tesla MRI简

This study aimed to investigate inflammatory edema after cerebral ischemia through 7.0T MRI and proton magnetic resonance spectroscopy（MRS）. All SD rats were randomly divided into sham operated group and middle cerebral artery occlusion（MCAO）-1 day,-3 day and-7 day groups. MRI scan of the brain was performed on a 7.0 Tesla MRI scanner. The volume of positive signals in the ischemic side was detected by using a T2 weighted spinecho multislice sequence; the changes in the height of water-peak were measured with point resolved spectroscopy（PRESS） sequences; cortical edema was detected by using wet-dry weight method; the degrees of nerve injury were evaluated by Bederson neurological score system; double-labeling immunofluorescence technique was used to explore the molecular mechanisms of post-ischemia cerebral edema. The results showed that high T2WI signals were observed in MCAO-1 day,-3 day and-7 day groups, and the water-peak height and water-peak area of MCAO groups were higher than those of sham operated group（P0.05）. Neurological score results were consistent with the degree of brain edema, and a large number of microglia accumulated in the ischemic cortex. Our results suggested that non-invasive MRI technology with the advantage of high spatial resolution and tissue resolution can comprehensively and dynamically observe inflammatory edema after cerebral ischemia from a three-dimensional space, and contribute to evaluation and treatments in clinic.     

INTRANASAL DELIVERY OF NERVE GROWTH FACTOR TO PROTECT THE CENTRAL NERVOUS SYSTEM AGAINST ACUTE CEREBRAL INFARCTION

Objective To confirmed reliability and feasibility of intranasal nerve growth factor (NGF) bypassing the blood-brain barrier and its potential neuroprotective effects on acute cerebral ischemia. Methods (1) To assay NGF concentrations in different brain regions after middle cerebral artery occlusion (MCAO).Rats were randomly divided into intranasal (IN) NGF, intravenous (IV) NGF, and untreated group (n = 4). The concentrations of NGF of different brain regions in the three groups after MCAO were measured by ELISA. (2) To observe neuroprotective action of NGF on focal cerebral ischemic damage. Rats were randomly assigned to 4 groups: IN vehicle, 1N NGF,1V vehicle, 1V NGF (n = 8). Treatment was initiated 30 minutes after onset of MCAO and given again 24 hours later. Three neurologic behavioral tests were performed 24 and 48 hours following onset of MCAO. Corrected infarct volumes were determined 48 hours after onset of MCAO. Results The olfactory bulb in IN NGF group obtained the highest concentration (3252 pg/g) of NGF among all regions, followed by the hippocampus. The NGF concentrations in the olfactory bulb and hippocampus in IN NGF group were markedly higher than that in 1V NGF and control groups. The infarct volume in IN NGF group was markedly reduced by 38.8% compared with IN vehicle group. IN NGF group vestibulum function markedly improved compared with IN vehicle group at 24 and 48 hours after onset of MCAO (P24h = 0.02 and P48h = 0.04, respectively), Conolmion Intranasal NGF could pass through the blood-brain barrier, reach the central nervous system, reduce infarct volume, and improve neurologic function in rats following MCAO. Intranasal delivery of NGF may be a promising treatment for stroke.     

Involvement of δ-and µ-opioid receptors in the delayed cerebral ischemic tolerance induced by repeated electroacupuncture preconditioning in rats

Background Preconditioning with repeated electroacupuncture （EA） could mimic ischemic preconditioning to induce cerebral ischemic tolerance in rats. The present study was designed to investigate whether mu（μ）-, delta（δ）- or kappa（κ）-opioid receptors are involved in the neuroprotecUon induced by repeated EA preconditioning. Methods The rats were pretreated with naltrindole （NTI）, nor-binaltorphimine （nor-BNI） or D-Phe-Cys-Tyr-D- Trp-Om-Thr-Pen-Thr-NH2 （CTOP）, which is a highly selective δ-, κ- or μ-opioid receptor antagonist respectively, before each EA preconditioning （30 minutes per day, 5 days）. Twenty-four hours after the last EA treatment, the middle cerebral artery occlusion （MCAO） was induced for 120 minutes. The brain infarct volume was determined with 2,3,5-tdphenyltetrazolium chloride staining at 24 hours after MCAO and compared with that in rats which only received EA preconditioning. In another experiment, the met-enkephalin-like immunoreactivity in rat brain was investigated by immunohistochemistry in both EA preconditioning and control rats. Results The EA preconditioning reduced brain infarct volume compared with the control rats （P=-0.000）. Administration of both NTI and CTOP attenuated the brain infarct volume reduction induced by EA preconditioning, presenting with larger infarct volume than that in the EA preconditioning rats （P〈0.001）. But nor-BNI administration did not block the infarct volume reduction induced by EA preconditioning, presenting with smaller infarct volume than the control group rats （P=-0.000）. The number of met-enkephalin-like immunoreactivity positive neurons in the EA preconditioning rats was more than that of the control rats （P=-0.000）. Conclusion Repeated EA preconditioning stimulates the release of enkephalins, which may bind 5- and p-opioid receptors to induce the tolerance against focal cerebral ischemia.     

Changes of evoked potential and expression of nestin in subventricular zones in rats after focal cerebral ischemia

Objective： To study the characteristics of latency of somatosensory evoked potential （SEP） and motor evoked potential （MEP） and the expression of nestin in subventricular zones （SVZ） after persistent focal cerebral ischemia in rats. Methods： The model of cerebral ischemia in rats was made by middle cerebral artery occlusion （MCAO）. All animals of ischemia were sacrificed after 12 h, 1 d, 3 d, 7 d, and 14 d to observe the changes of latency of SEP and MEP and to detect the expression of nestin, with an immunohistochemical approach. Results： The latencies of P1 （positive wave 1）, N1 （negative wave 1） and P2 （positive wave 2） in SEP were significantly prolonged after MCAO. The latencies of N1 and N2 waves in MEP were postponed gradually and no statistical difference of latency of N1 wave was found in rats at 7 d and 14 d after MCAO. The expression of nestin increased at 12 h, and showed a significant augmentation at 3 d and peaked at 7 d, then declined slightly at 14 d after MCAO. Conclusion： The cerebral ischemia prolonged the latency of EP waves and the expression of nestin was up-regulated and reached the peak at 7 d, showing the ischemia induced the proliferation of nervous stem cells. The SEP and MEP may evaluate the proliferation in SVZ after brain ischemia.     

Acupuncture Increases Bone Strength by Improving Mass and Structure in Established Osteoporosis after Ovariectomy in Rats

The effects of acupuncture on bone biomechanical properties and histomorphometry in ovariectomized （OVX） rats were studied. Twenty-four 8-week-old female Sprague-Dawley rats were randomly assigned to three groups： sham, model and acupuncture, Rats in the model and acupuncture groups were ovariectomized, while those in the sham group underwent a sham operation. All rats were anesthetized and fastened for 15 minutes, and for the acupuncture group, needling on Pishu （BL20） and Shenshu （BL23） was performed. Blood and urine were collected to measure serum osteocalcin （OC） and urinary calcium, phosphorus or deoxypyridinoline （Dpd）. After 16 weeks of treatment, all the rats were killed and their tibiae and femora were removed. The tibiae were used for analyses of bone histomorphometry and the femora for a three-point bending test. Acupuncture gave significant protection against ovariectomy-caused decline on femoral strength in the mechanical test, increased the trabecular bone volume and thickness, lowered the trabecular separation of tibiae and restricted the excretion of phosphorus and Dpd, while promoting the concentrations of serum osteocalcin as compared with model rats. These results seemed to indicate that acupuncture on the points of Pishu （BL20） and Shenshu （BL23） not only promoted the bone formation but also suppressed the bone resorption induced by OVX in osteoporotic rats, which suggests that it would be a potentially useful and convenient method in preventing osteoporosis.     

Effect of Transcranial Magnetic Stimulation on the Expression of c-Fos and Brain-derived Neurotrophic Factor of the Cerebral Cortex in Rats with Cerebral Infarct

The effect of transcranial magnetic stimulation （TMS） on the neurological functional recovery and expression of c-Fos and brain-derived neurotrophic factor （BDNF） of the cerebral cortex in rats with cerebral infarction was investigated. Cerebral infarction models were established by using left middle cerebral artery occlusion （MCAO） and were randomly divided into a model group （n=40） and a TMS group （n=40）. TMS treatment （2 times per day, 30 pulses per time） with a frequency of 0.5 Hz and magnetic field intensity of 1.33 Tesla was carried out in TMS group after MCAO. Modified neurological severity score （NSS） were recorded before and 1, 7, 14, 21, and 28 day（s） after MCAO. The expression of c-Fos and BDNF was immunohistochemically detected 1, 7, 14, 21, and 28 day（s） after infarction respectively. Our results showed that a significant recovery of NSS （P〈0.05） was found in animals treated by TMS on day 7, 14, 21, and 28 as compared with the animals in the model group. The positive expression of c-Fos and BDNF was detected in the cortex surrounding the infarction areas, while the expression of c-Fos and BDNF increased significantly in TMS treatment group in comparison with those in model group 7, 14, 21, and 28 days （P〈0.05） and 7 14, 21 days （P〈0.01） after infarction, respectively. It is concluded that TMS has therapeutic effect on cerebral infarction and this may have something to do with TMS＇s ability to promote the expression of c-Fos and BDNF of the cerebral cortex in rats with cerebral infarction.     

Expression of Arginyl-tRNA Synthetase in Rats with Focal Cerebral Ischemia

Aminoacyl-tRNA syntheses （AARS） can catalyze the adenosine triphosphate （ATP）-dependent acylation of their cognate tRNA（s） with a specific amino acid. They can be seen as an index to reflect the energy metabolic rate of ischemic brain cells in ischemic penumbra. This study ex- amined the relationship between arginyl-tRNA synthetase （ArgRS）, one of the AARS, and cerebral ischemia in rats. The model of middle cerebral artery occlusion （MCAO） was established in rats. The expression levels of ArgRS protein and mRNA were detected in rat brain tissues at different time points following MCAO by Western blotting and RT-PCR, respectively. The results showed that the MCAO model was successfully established. Western blotting and RT-PCR analysis revealed that the ArgRS protein and mRNA were expressed in brain cells in both ischemic and normal penumbra tissues. The expression levels of ArgRS protein and mRNA peaked at 6 h after MCAO and decreased gradually. At 24 h, the expression levels of ArgRs protein and mRNA in ischemic penumbral tissues were lower than those in normal tissues. The expression levels of ArgRS mRNA and protein in ischemic penumbra var- ied with ischemic time, suggesting that the energy metabolism of brain cells in penumbra changed dy- namically after ischemia to ensure the endogenous self-protection of the body. The brain oxygen supply should be improved as soon as possible, especially within 6-12 h after ischemia, so as to meet the de- mand for energy metabolism in ischemic penumbra and make sure the cell structure remains stable.     

抗骨疏不同提取物大鼠模型骨质疏松症的作用及其潜在机制

Objective： To investigate the preventive effects and possible underlying mechanism of different extracts of Kanggushu （抗骨疏） on osteoporosis in ovariectomized rats. Methods： One hundred and sixty- five female SD rats were divided into 11 groups： control, sham, model, Xianling Gubao Capsule （仙灵骨葆胶囊）, nilestriol, Kanggushu aqueous extract high-, medium-, and low-dose and suet extract high-, medium-, and low-dose groups. The osteoporosis model was made by ovariectomizing the rats. The latter 8 groups were administered intragastricly with Xianling Gubao Capsule, nilestriol, Kanggushu aqueous extract and suet extract for 12 weeks, respectively, while the other 3 groups were administered orally saline. The whole body bone mineral density, bone mineral content, organ coefficient of uterus, serum estradiol and alkaline phosphatase contents, blood calcium, phosphorus, interleukin 6 and bone Gla-protein levels after treatment were monitored. Additionally, three-point bending test of femur, HE staining, and scanning electron microscope were performed to explore the pharmacodynamics and underlying mechanisms. Results： In comparison with ovariectomized rats of model group, Kanggushu aqueous extract high-dose resulted in an increased bone mineral density, bone mineral content and organ coefficient of uterus, improved estradiol level, and improved maximum load and structural stiffness （P〈0.05 or P〈0.01）. Two-dimensional and thrae-dimensional trabecular structure was also observed under HE staining and scanning electron microscopy, and the number and thickness of trabecular bone in Kanggushu aqueous extract high-dose group was significantly increased compared to the model group, while the lipid droplets in bone marrow cavity were significantly less. However, there were no significant differences in blood calcium, total serum alkaline phosphatase and bone Gla protein among different treatment groups. Overall, the osteoprotective effects of Kanggushu aqueous extract were comparable to those of nilestriol and were significantly more effective than those of Xianling Gubao Capsule. Conclusion： The preventive effects of Kanggushu aqueous extract might be partly due to the increased estradiol level, accelerated restoration of bone trabecular reticulate structure, and accordingly increased bone mineral density in osteoporosis rats.     

Study on preventive effect of jiangu granule on osteoporosis in ovariectomized rats

Objective:Toinvestigatethepreventiveef fectofJiangu granule(JGG)onexperimentalprimaryosteoporosistypeⅠ.Methods:Osteo porosismodelwasestablishedthroughovaryre sectionoffemalerats.Bonemineraldensity(BMD)wasmeasuredwithdoubleenergyX rayabsorptiometry.Levelo…     

Effect of acupuncture at acupoint of Shenshu (BL 23) on the bones of ovariectomized rats

OBJECTIVE

To investigate the effect of acupuncture at acupoint of Shenshu (BL 23) in ovariectomized rats and sham-operated rats.

Neutroprotective efficacy of sodium tanshinone B on hippocampus neuron in a rat model of focal cerebral ischemia

<正>Objective:To investigate the protective effects of sodium tanshinone B(STB) on brain damage following focal ischemia-reperfusion(l/R) injury through interfering with N-methyl-D-aspartic acid receptor(NMDAR) and excitatory and inhibitory amino acids,and evaluate the potential mechanisms of the neuroprotective activity of STB.Methods:Transient forebrain ischemia was induced by middle cerebral artery occlusion(MCAO).The rats were randomized into a sham operated group,a model group(l/R) and three STB different dose groups.Rats were pretreated with STB at the doses of 4,8,16 mg/kg(STB_1,STB_2,STB_3) for 3 days before MCAO.The expression of NMDAR1 was detected by immunohistochemistry and Western blotting. The concentrations of glutamate and y -aminobutyric acid(GABA) were analyzed using high performance liquid chromatography.Results:STB treatment reduced neurological defect scores,cerebral infarction volume and brain water content.The levels of NMDAR1 were significantly higher in the l/R and STB,groups than that of the sham and the STB_3 groups(P0.01).Optical density of NMDAR1 was significantly increased in cornu ammonis(CA)1 region of the l/R group(P0.05).STB treatment reduced NMDAR1 optical density in the CA1 region(P0.01).The levels of glutamate were significantly lower in the hippocampus in the STB_3 group than that of the l/R,STB,and STB_2 groups(P0.01).Conclusion:Preconditioning with STB appears to be a simple and promising strategy to reduce or even prevent cerebral l/R injury and has potential for future clinical application.     

Clinical biochemical observation on the treatment of postmenopausal osteoporosis with Gusong II

Objective: To assess the effect of Gusong II in treating postmenopausal osteoporosis and try to reveal its mechanism.Methods: Thirty-five women, 49 to 62 years old, with 5.6 years menopause, were treated as the subjects and followed-up for 6 months, 12 months or 24 months. The course of treatment ranged from 6 months to 25 months with an average of 14 months. Some related biochemical parameters, such as serum estradiol (E₂), testosterone (T), the ratio of fasting urinary calcium to creatinine (Ca/Cr), the ratio of 24h urinary calcium to creatinine (24h Ca/Cr), 24h urinary E₂, and 24h urinary hydroxyproline (HOP) were measured before and after treatment.Results: After 6~9 months or 1~2 years of treatment, the levels of serum E₂ and 24h urinary E₂ were raised significantly, while the levels of serum T and urinary Ca/Cr fell evidently after one or two year’s treatment. The levels of serum E₂ and 24h urinary E₂ were significantly correlated with the levels of fasting urinary Ca/Cr, 24h urinary Ca/Cr, serum T, and 24h urinary HOP respectively.Conclusions: Gusong-II can elevate the serum E₂ levels of postmenopausal women, reduce the discharge of urinary calcium and HOP, and inhibit bone resorption. The rise of the serum E₂ level might be correlated with the conversion of T to E₂.     

玉屏风散对慢性支气管炎大鼠的抗炎及免疫调节作用

Objective: To investigate the anti-inflammatory and immunoregulatory effects of Yupingfeng (玉屏风, YPF) Powder and its components in rats. Methods: A rat chronic bronchitis (CB) model was developed using lipopolysaccharide (LPS) combined with bacillus Calmette Guerin (BCG). YPF, simple recipe Astragalus membranaceus (Fisch.) Bge (AM) and Astragalus membranaceus (Fisch.) Bge plus rhizome of Atractylodes macrocephala Koidz (AM+RA) decoction were administered (intragastric administration, once a day for 21 days) to rats, to prevent and treat CB. Immunoregulatory and anti-inflammatory effects of YPF, AM and AM+RA were tested by serum pharmacology in vitro on splenic lymphocytes of normal rats and alveolar macrophages of CB rats. Results: Inflammation in the pulmonary tissue and the bronchus of CB rats was significantly reduced in the YPF-treatment groups, AM and AM+RA groups demonstrating the efficacy of YPF. Serum samples collected at different times from rats after administration of YPF, AM and AM+RA demonstrated increased proliferation of splenic lymphocytes with area under the effect curve (AUE) of 552.6%, 336.3% and 452.0%, respectively. Treatment of alveolar macrophages with serum samples in YPF, AM or AM+RA group inhibited interleukin-8 (IL-8) in the cell culture media, and the effect was much better in the YPF group compared with AM or AM+RA group, with a higher maximal effect (Emax, P<0.05) and larger AUE (P<0.01 and P<0.05). Moreover, serum from rats treated with AM or AM+RA had similar efficacy, while the efficiency was lower than that treated with YPF. Conclusion : YPF demonstrated anti-inflammatory and immunoregulatory effects in a rat model of CB, and time-dependent relationships were demonstrated in vitro.     

雌激素对偏头痛大鼠三叉神经节内降钙素相关基因肽mRNA表达的影响

目的 :研究雌激素对偏头痛大鼠三叉神经节内降钙素相关基因肽 (CGRP)mRNA表达的影响 .方法 :成年雌性SD大鼠去卵巢后分为 3组 :去卵巢组 (OVX)、低雌激素替代组 (OVX +LDET)及高雌激素替代组 (OVX +HDET) ,随后各组内部分大鼠以皮下注射硝酸甘油 (NTG)复制偏头痛模型 ,采用RT PCR方法观察三叉神经节内CGRPmRNA表达的变化 .结果 :去卵巢组与雌激素替代组比较 ,三叉神经节内CGRPmRNA表达增强 (P <0 .0 5 ) ,去卵巢及低雌激素替代组硝酸甘油造模后大鼠三叉神经节内CGRPmRNA与造模前比较表达增强 (P <0 .0 5 ) ,而高雌激素替代组造模前后无明显变化 (P >0 .0 5 ) .结论 :实验性偏头痛大鼠三叉神经节内CGRPmRNA的过度表达可能是偏头痛发作的分子机制之一 ,雌激素通过调控三叉神经节内CGRPmRNA的表达 ,可能参与女性偏头痛的发生     

Hormone-like activities of Kuntai capsule in the uteri of ovariectomized rats and immature rabbits

OBJECTIVE

To investigate the hormone-like activities of Kuntai capsule (KTC) in the uteri of ovariectomized rats and immature rabbits.

去卵巢和抗骨质疏松干预后大鼠尿脱氧吡啶啉的变化

目的观察大鼠去卵巢及予以不同的干预后尿脱氧吡啶啉/肌酐比率(urinarydeoxypyridinolinecrosslink/creatinine,UDpd/Cr)的变化,了解其在骨质疏松的诊断和治疗中的应用价值。方法将40只Wister大鼠于6月龄时行去卵巢手术(OVX,n=32)和假手术(Sham,n=8)。手术后6周分别予以不同的干预如下:OVX组(Ovx,n=8)和Sham组不做干预;雌激素干预组(O+E,n=9)予以17β雌二醇20μg/(kg·d),皮下注射;英卡膦酸钠干预组(O+C,n=7)予以Cimadronate0.2mg/(kg·d),经胃肠给药。手术后14周将大鼠处死,处死前收集24h尿液和血液测定血清及尿液中生化指标;处死后留取右侧股骨标本,进行股骨外周定量CT骨密度及股骨远端凹入实验生物力学测定。结果OVX组大鼠的骨吸收指标UDpd/Cr比Sham组增加了133.3%(P<0.01),OVX组大鼠尿钙/肌酐比率亦有增加,但与Sham组相比差异无显著性(P>0.05)。雌激素和英卡膦酸钠干预组UDpd/Cr比OVX组显著降低,分别减少了54.6%和51.8%(P均<0.01),并与Sham组接近。UDpd/Cr与骨量、骨密度及骨生物力学等指标的相关分析表明,其与以上指标之间存在显著负相关。结论UDpd/Cr是敏感的骨吸收指标,在OVX和予以雌激素、二膦酸盐干预后均有显著变化,其改变与骨密度及骨骼力学特性的改变密切相关。