Effects of estradiol benzoate on proliferation and differentiation of in vitro cultured hippocampal neural stem cells / 中华老年医学杂志

Objective To explore the potential mechanism of action of estradiol benzoate on proliferation and differentiation of rat hippocampal neural stem cells.Methods Hippocampus tissues from SD rats were dissociated and plated into culture flasks.Neural stem cells were identified by immunofluorescence against Nestin.After treatment with different concentration of estradiol benzoate (10-10,10-9,10-8,10-7,10-6 mol/L),the proliferation and differentiation of neural stem cells were assessed by MTT assay and the proportion of neurons and astrocytes in differentiated cells was assessed by flow cytometry.Western blot assay was performed to detect the expression levels of protein in neural stem cells.Results (1) The neural stem cells formed neurospheres and grew in floating,and were Nestin-positive.Estrogen receptors,including ERα and ERβ,were expressed in neural stem cells;(2)After treatment with 10-8 mol/L estradiol benzoate versus in the control group,the cell viability was significantly increased (t =5.36,P =0.003),and the ratio of neuron-specific nuclear protein(NeuN)-positive neuron cells in total cells was significantly increased (t =4.32,P =0.02),but the ratio of glial fibers acidic protein(GFAP)-positive cell in total cells was decreased(t=4.65,P=0.008).The expression levels of GDNF,GFRal and Ret proteins were enhanced in the estradiol benzoate group compared with the control group.Conclusions Appropriate concentration of estradiol benzoate promotes proliferation and differentiation ofin vitrocultured hippocampal neural stem cells and elevates the ratio between neurons and glial cells differentiated from neural stem cells.

Effect of atorvastatin on brain edema and its mechanism in rats after focal cerebral ischemia / 中华神经医学杂志

Objective To observe the dynamic changes of brain edema,tumor necrosis factor-α (TNF-α),aquaporin-4 (AQP-4) in focal cerebral ischemia rats and investigate the effect of atorvastatin on the above-mentioned factors.Methods One hundred and eight male SD rats were randomly divided into sham-operated group,model group and treatment group (n=36);furthermore,each group was divided into three subgroups:1 d group,3 d group and 7 d group (n=12).Focal cerebral ischemia models in model group and treatment group were induced by left middle cerebral artery occlusion,while rats in the sham-operated group were only separated from blood vessels with no ligation and no inserted line.After succeeded models,the treatment group received gavage administration of atorvastatin (4 mg/ [kg·d]),others were given the same dosage of normal saline at the same time.The brain edema was calculated by measuring brain water content via dry-wet method,and the mRNA expressions of TNF-α and AQP-4 were detected by real time-PCR;Western blotting was employed to detect the AQP-4 protein expression.Results At the same observation time,as compared with those in the sham-operated group,the brain water content,TNF-α mRNA expression and A QP-4 mRNA and protein expressions in the model group and treatment group markedly increased (P＜0.05);as compared with the model group,treatment group had statistically decreased brain water content,TNF-α mRNA expression and AQP-4 mRNA and protein expressions (P＜0.05).Conclusion The brain edema of focal cerebral ischemia rats may be positively correlated to AQP-4 mRNA and protein expressions induced by TNF-α;atorvastatin probably down-regulates AQP-4 mRNA and protein cxpressions by inhibiting TNF-α to alleviate brain edema and protect brain tissues.

Research on adipose-derived stem cells differentiation induced by supernatant liquid from cerebral infarction tissue in rats / 中华老年医学杂志

Objective To study adipose-derived stem cells (ADSCs)differentiation into endothelial cells and smooth muscle cells by induction with supernatant liquid from cerebral infarction tissue in rats.Methods The ADSCs were obtained from retroperitoneal adipose tissue of rats and identified by flow cytometry technology.The normal brain tissues and the infarcted cerebral tissue obtained from rats with middle cerebral artery occlusion were used to induce ADSCs differentiation,and no intervention group was as a control.The mRNA expression level of von willebrand factor (vWF),α-smooth muscle actin (α-SMA) and smooth muscle myosin heavy chain (SM-MHC) in cells after induction were detected using reverse transcriptase-polymerase chain reaction.Immunofluorescence were used to identify the expression of endothelial cells markers and smooth muscle cell markers,and positive expression cell was detected by fluorescence microscope.Results On the ADSCs surfaces,the high expressed-positive antigens included CD90 (96.7％),CD29 (84.4％),CD44 (98.9％),and the low expressed negative antigens included CD45 (6.5％),CD34 (7.4％) and CD31 (3.6％).Compared with no intervention group and normal brain tissue of supernatant liquid group,the cerebral infarction tissue of supernatant liquid-induced group showed the increased mRNA expression level of vWF,α-SMA and SM-MHC(F=5.962,6.756,6.144,P=0.001,0.004,0.003),and showed that the immunofluorescence indicated-cell expression level of vWF,α-SMA and SM-MHC was much more increased (all P ＜ 0.05).Conclusions Under the induction by supernatant liquid of cerebral infarction tissue,ADSCs highly expresses the markers of endothelial cells and smooth muscle cells.This suggests that the cerebral infarction tissue of supernatant liquid-induced ADSCs have a tendency to differentiate into endothelial cells and smooth muscle cells.

Effect of atorvastatin on angiogenesis of brain tissues in focal cerebral ischemia rats / 中华神经医学杂志

Objective To investigate the effect ofatorvastatin on angiogenesis of brain tissues in focal cerebral ischemia rats,and explore the corresponding mechanism.Methods Ninety male SD rats were randomly divided into sham-operated group,model group and treatment group (n=30);and then,each group was divided into three subgroups:1 d group,3 d group and 7 d group (n=10).The focal cerebral ischemia models were induced by middle cerebral artery occlusion (MCAO);3 h after MCAO,rats in the treatment group received a gavage of atorvastatin 4 mg/(kg.d),and others were given the same amount of normal saline at corresponding time.The nerve function defects were estimated at 3 h after MCAO and before being killed;the protein expressions of vascular endothelial cell markers CD105 and glucose-regulated protein 78 (GRP78) were analyzed by immunohistochemistry;the vascular endothelial growth factor (VEGF) mRNA expression was analyzed by real time-PCR;the caspase-12 protein expression was analyzed by Western blotting.Results (1) Nerve function defects scores:there was no significantly statistical difference between model group and treatment group at 3 h after MCAO (P＞0.05),but statistical differences at different time points before being killed were noted (P＜0.05).(2) Microvessel density (MVD):at all time points,that of model group was increased as compared with that of sham-operated group,that of treatment group was increased as compared with that of model group,and the differences were all statistically significant (P＜0.05).(3)The VEGF rmRNA expression gradually increased over time;at all time points,the VEGF mRNA expression of model group was significantly higher than that in the sham-operated group,and that of treatment group was significantly higher than that in the model group (P＜0.05).(4)The GRP78 and caspase-12 expressions were gradually decreased over time;at all time points,the GRP78/BiP and caspase-12 expressions in the model group were significantly higher than those in the sham-operated group;those in the treatment group were statistically lower than those in the model group,but obviously higher than those in the sham-operated group (P＜0.05).Conclusion The angiogenesis of brain tissues in MCAO rats is obvious,and atorvastatin can enhance this effect;the mechanism may be that atorvastatin can weaken the endoplasmic reticulum stress,and then,reduce the apoptosis of endothelial cells.

Study on chemical constituents of Scutelliaria regeliana / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents in ethyl acetate fraction from the root of Scutelliaria regeliana.</p><p><b>METHOD</b>The compounds were isolated by silica gel column chromatography and HPLC, and their structures were elucidated by means of spectral analyses.</p><p><b>RESULT</b>23 compounds were isolated and identified.</p><p><b>CONCLUSION</b>Compound 1 is new, named as scutellariae flavonol, and the others were isolated from S. regeliana for the first time.</p>

Clinical presentations of smog disease / 中国基层医药

Objective To investigate the features of smog disease.Methods The clinical manifestations and inducements of smog disease patients were comprehensively analyzed by using clinical statistical methods.Results Smog disease mainly invaded young people.Smog disease was found mostly acute or subacute in onset.21 cases invaded internal carotid arterial system among these 30 patients,including infarction and cerebral hemorrhage;6 patients showed vertebral arterial signs,and 3 cases invaded both the two systems.Brain CT or MRI only showed the position of neurological disorder;The final diagnosis was made by DSA.Conclusion Smog disease could invade both the two systems of intracerebral artery,DSA is the best method to diagnose smog disease.

Platelet activation and brain protection of protein hydrolysate injection for patients with acute stroke / 中国组织工程研究

BACKGROUND: The blood levels of specific markers of platelet activation, such as platelet granule membrane protein (GMP-140) and tumor necrosis factor alpha (TNF-α) are very low in healthy individuals, while the plasma levels of them in patients with cerebral infarction increase. Is the protection of brain hydrolysate injection correlated with the phenomenon?OBJECTIVE: In this study, the plasma levels of GMP-140 and TNF-α in patients with acute stroke were measured, the brain protection of brain protein hydrolysate injection on patients with ischemic stroke were investigated, and were compared with the therapeutic effect of compound danshen injection.DESIGN: It was designed for case study.SETTING: This study was conducted at the Medical Department of the General Hospital of Pingdingshan Coal Co. Ltd and the Neurology Department of the Second Hospital Affiliated to Zhengzhou University.PARTICIPANTS: From January 2001 to October 2003, 144 inpatients with hypertension and acute stroke in the Medicine Department of the General Hospital of Pingdingshan Coal Co. Ltd were selected and divided into 2 groups, as experiment group containing 72 cases, 47 males and 25females, with an age from 42 to 90 and in average of (69±11) years old and control group containing 72 cases, 49 males and 23 females, with an age from 37 to 85 and in average of (68±10) years old.METHODS: All the patients in these two groups underwent oxygen inhalation therapy, antihypeetensive therapy, dehydration therapy and anticoagulation therapy. Patients in control group were coadministered 500 mL compound salvia miltiorrhiza injection QD once a day, with a 14-day course of therapy. Patients in experiment group were treated with 500 mL compound salvia miltiorrhiza injection QD and 20 mL protein hydrolysate European stroke scale (ESS), from 0 (worst possible health status) to 100(best possible health status), were used to evaluate the recovery status of (from 24 hours to 72 hours) and 3 weeks post-treatment, 5 mL blood samples were obtained from antecubital veins, then plasma levels of GMP-140and TNF-α were quantified using an RIA (radioimmunoassay) and the changes in neural function before and after brain protein hydrolysate injection were evaluated.MAIN OUTCOME MEASURES: Before treatment and at 3 weeks postwere quantified using an RIA.RESULTS: All the 144 patients entered the statistical analysis procedure.ESS were significantly higher than the pre-treatment scores [Experiment groups: 79.95±18.64 and 59.65±19.87; Control group: (74.66±15.88) and (61.25±18.68), (t=2.678-4.351, P ＜ 0.01). The post-treatment scores of ESS in experiment group were higher than those in control group (t=2.016-2.158,groups, the post-treatment outcomes were significantly lower than the pre-treatment outcomes [Experiment group: (22.12±9.52) μg/L and (50.41±22.35) μg/L, (1.05±0.24) μg/L and (1.62±0.50) μg/L; Control group: (26.66±8.22) μg/L and (48.63±21.54) μg/L, (1.35±0.44) μg/L and (1.66±0.48) μg/L; (t=2.678-4.351, P ＜ 0.001)]. And the post-treatment levels of the two markers were lower in experiment group than those in control group (t=2.016-2.158, P ＜ 0.05).CONCLUSION: Brain protein hydrolysate injection can significantly decrease the plasma levels of GMP-140 and TNF-α in patients with acute stroke, and it is capable of increasing the ESS scores and improving the impaired neural functions greatly.

Puerperal practice pattern in a rural area of north China / 北京大学学报(医学版)

Objective:To describe women's behavioral patterns in the period of “doing the month”(confinement in childbirth)and to determine the degree of differences between “doing the month” practices and daily behaviors.Methods:A retrospective survey was conducted with a random sample of 1 813 women who were interviewed using a questionnaire undertaken in May of 2004.Results:The results showed that the majority of women adhered to the traditional customs.While “doing the month”,more than 90% of women did not eat cold,hard,or sour food;more than 90% of the women did not wash their hair or body at all,and more than 70% of women did not brush their teeth or wash their!feet.As compared to what women normally do,most women drastically changed their practices in eating,sanitation,dress,activity,doing chores,and so on,during their “doing the month”.Further analysis indicated that women's age had no effect on the change of practice patterns and that women's education had an inconsequential effect.Conclusion:The traditional “doing the month” practices are still very popular in the rural areas of China.Birth belief,illness concept and traditional village culture are the main factors for the pattern of “doing the month”.