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Objective To investigate the effect of chaihu-shugan-san ( CSS) on the behaviors and Notch1 signal pathway in depression model rats. Methods Thirty-two SD rats with similar behavioral scores were divided into control group (CON),model group (CUMS),positive control group (FLU) and interven-tion group (CSS).The depression model was established by stimulating with chronic unpredictable mild stress (CUMS),and the behaviors evaluation was assessed by sugar water consumption and forced depression.Im-munofluorescence was used to detect the proliferation of hippocampus neurons in rats,at the same time,real- time PCR and Western blot were used to detect the mRNA and protein expression levels of each factor (Notch1,Hes1,Hes5 and Jagged1) of Notch1 signal pathway respectively. Results Compared with CON group,the percentage of sugar water preference and swimming length of rats decreased significantly in CUMS group (P＜0.05 and P＜0.01).Compared with CUMS group,the percentage of sugar water preference and swimming length of rats increased significantly in CSS group(P＜0.05).Compared with CON group,there was a significant increase in the inactivity length of rats between CUMS group,FLU group and CSS group,and the difference was statistically significant (P＜0.01).Compared with CUMS group,the swimming length of rats in CSS group was significantly reduced,and the difference was statistically significant (P＜0.01).Compared with CON group((750.00±27.51)/mm2),the number of BrdU positive cells in the substratum or granulocyte lay-er of the hippocampus dentate gyrus of rats in CUMS group ((338.75±29.61)/mm2),FLU group ((545.00 ±17.73)/mm2) and CSS group ((529.38±13.74)/mm2) was significantly reduced(P＜0.01).Compared with CUMS group ((338.75±29.61)/mm2),there was a significant increase in the number of BrdU positive cells in the substratum or granulocyte layer of the hippocampus dentate gyrus of rats in CSS group ((529.38 ±13.74)/mm2),and the difference was statistically significant (P＜0.01).Compared with CON group,the mRNA and protein expression levels of Notch1,Hes1,Hes5,and Jagged1 in the hippocampus of rats in CUMS group were significantly reduced(P＜0.05 or P＜0.01).Compared with CUMS group,the mRNA and protein ex-pression levels of Notch1,Hes1,Hes5 and Jagged1 in the hippocampus of rats in FLU group and CSS group were significantly increased,and the differences were statistically significant (P＜0.05 or P＜0.01). Conclu-sion Notch1 signal pathway may be related to the obstacle during the hippocampus nerve regenerating in the model rat under chronic unpredictable mild stress.CSS may play an anti-depressant role by regulating Notchl to improve hippocampus nerve regeneration.
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To explore the presence of informative protein biomarkers in the salivary proteome of breast cancer patients with thick white or thick yellow tongue fur.
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Objective To observe the effect of aqueous extract of Broussonetia papyrifera ( L. ) Vent on the ability of space learning and memory in the rats with Alzheimer' s disease (AD) induced by Aβ 25-35 and Dgalactose and to explore the mechanisms underlying those improvements. Methods The animal model of AD was established by Aβ 25-35 stereotactic injection into the hippocampus of rats in 5 minutes,while long-term intraperitoneal injection with D-gal. After the injection of Aβ25-35,rats were treated with aqueous extract of Broussonetia papyrifera ( L. ) Vent for the next 30 days. Morris water maze with computer system and the spatial exploration experiments were used to assess the behavior performances of the rats. Immunohistochemical technique was used to detect the expression of BiP, PERK and CHOP. Results The ability of space learning and memory of rats complex model of Alzheimer's disease induced by Aβ25-35 and D-gal was damaged,while escape latency was (20.90± 9.16 ) s,and the proportion of original platform quadrant was ( 11.05 ± 4.43 ) %. The expression level of Bip was reduced ,while the mean gray was ( 139.71 ± 3.47 ). The expression level of PERK and CHOP was increased,while the mean gray were (97.96 ± 5.97 ), ( 110.93±4.91 )separately. The escape latency of rats in the aqueous extract of Broussonetia papyrifera ( L. ) Vent treated groups was ( 5.41 ± 3.47 ) s and shorter than the model group,while the proportion of original platform quadrant was (48.28 ± 7.03 )% and higher than the model group.The expression level of Bip in the treated group was higher than the model group, while the mean gray were ( 121.17 ±4.76). The expression level of PERK and CHOP in the treated group was lower than the model group,while the mean gray were ( 122.11 ± 4.73 ), ( 123.34 ± 7.73 ) separately. Significant differences were observed between model group and aqueous extract of Broussonetia papyrifera ( L. ) Vent treated groups (P＜ 0. 05 ～ 0. 01 ).Conclusion Aqueous extract of Broussonetia papyrifera ( L. ) Vent can improve learning and memory disorders of the model rats induced by Aβ25-35 and D-galactose. ER (endoplasmic reticulum) stress and correlated apoptosis pathway might be involved in the underlying mechanisms.
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BACKGROUND: Current methods of stem cell separation are mainly based on their cell markers.A method for stem cells separation which is not based on cell markers developed in recent years,that is fluorescence activated cell sorting method,has been applied for stem cells and mature cells separation.OBJECTIVE: To isolate side population cells from newborn mice small intestinal mucosa,and to investigate the feasibility of constructing the murine intestinal stem cell population by fluorescence activated cell sorting.METHODS: Small intestine mucosa organoids of mice were isolated and dissociated into single cells.The side population cells were stained with Hoechst 33342 and propidium iodide,then sorted using fluorescence activated cell sorting.Total RNA and protein were purified from sorted fractions to detect Musashi-1 expressions by RT-PCR and Western-blotting.RESULTS AND CONCLUSION: Single cell suspension from mouse small intestine mucosa contained a viable population of cells,which showed the side population phenotype and were sensitive to verapamil.These cells were enriched for Musashi-1 mRNA and MSI-1 protein expression.Results demonstrated that the side population fraction separated from mice intestinal mucosa is enriched for intestinal stem cells,the murine intestinal stem cell population can be successfully constructed with fluorescence activated cell sorting.
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Objective To explore mRNA and protein expression of PAl-1 in kidney tissue of mice with IgA nephropathy (IgAN) and the effects of YiShen Decoction. Methods The IgAN model was built by the method of oral intake of bovine serum albumin(BSA)together with the injection of staphylococcus enterotoxin B(SEB)through caudal vein. The mRNA expression of PAI-1 was measured by fluorescent quantitative polymerase chain reaction (FQ-PER)and the expression of PAI-1 protein was measured by immunohistochemistry. Results No significant difference of mRNA and protein expression of PAI-1 was found between the low concentration and high concentration Yishen Decoction group. But the secretion and mRNA expression of PAI-1 in the low and hiigh concentration Yishen Decoction group was decreased more than that in the IgAN model group. Conclusion The abnormal expression of PAI-1 mRNA and protein played an important role in the onset and development of IgAN. The TCM Yishen Decoction could reduce the abnormal expression of the mRNA and protein of PAI-1 in kidney tissue of mice with IgA nephropathy.
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Objective To investigate the microbiostatic mechanism of denuded tongue coating, Proteomic methods were used to find differential expression proteins in denuded tongue. Methods The total protein of normal or denuded tongue coating was separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). The differential expression protein was analyzed by using ImageMaster 2D analysis software. Image analysis software was used for comparative analysis to find differential expression proteins; Mass spectrometry (mass spectrometry) was applied to get the corresponding peptide fingerprint (peptide mass finger print_ PMF). Search the database to identify differences in protein.Results Establish two-dimensional gel electrophoresis for normal and denuded tongue; Segregation analysis on four groups of proteome of tongue coating showed that gel could be detected in the protein spots at 1082=t=105 and 1143±140 respectively; Compared with normal group, there are eight points of protein expression being regulated upward and seven points down.Altogether 9 differences in protein expression were identified. Conclusion The sound resolution and repeatability Two-dimensional electrophoresis silver staining pattern were established; Some differences in protein expression related to denuded tongue were identified.
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Objective To investigate the effect and mechanism of invigorating spleen,eliminating stasis, heat-clearing drugs(milk veteh,root of red rooted salvia,Chinese goldthread)on quality of ulcer healing (QOUH).Methods The rat models of chronic gastric ulcer was induced by acetic acid,the effects of QOUH of milk veteh,root of red rooted salvia and Chinese goldthread on the model of gastric Ulcer were observed, the expression of EGFR mRNA of mucosa were measured by in situ hybridization.Results Milk veteh,root of red rooted salvia and Chinese goldthread could improve the injury of gastric mucosa.Milk veteh,root of red rooted salvia increased the expression of EGFR mRNA in the tissue around peptic ulcer(PU). Conclusion Invigorating spleen,eliminating stasis,heat-clearing drugs can improve QOUH,it is one of possible mechanisms that the drugs increased the expression of EGFR mRNA in the tissue around PU.
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BACKGROUND: Tiantai I consists of gastrodia, Chinese angelica root, areca seed. It has been considered as the roles of invigorating the liver, nourishing marrow, heightening the intelligence and causing resuscitation. OBJECTIVE: To observe the effects of Tiantai Ⅰ on the abilities of learn ing and memory and the activity of central cholinergic system in mice withspontaneous Alzheimer disease. DESIGN: A randomized control observation. SETTING: Shenzhen Institute of Integrated Chinese and Western Medicine. MATERIALS: Male Kunming mice of 13 months old were raised to 21 months old, of which 52 with spontaneous Alzheimer disease werescreened. They were randomly divided into blank control group, westerndrug control group, Tiantai Ⅰ 6.80 and 20.41 g/kg groups, 13 mice in eachgroup. Another 13 aged mice with normal learning and memory abilitieswere selected as the normal control group at the same time. METHODS: Mice in the western drug control group were treated with0.6 mg/Kg Hydergine, those in the Tiantai Ⅰ 6.80 and 20.41 g/kg groupswere given intragastric administration of Tiantai Ⅰ of 6.80 and 20.41 g/kg, respectively, and those in the normal control group and blank control groupwere given double distilled water of the same volume. The learning andmemory results were examined by the step-down test. Freezing sections ofbrain tissue were prepared, acetylcholinesterase (AChE) fiber was showedaccording to the Hedreen method, and choline acetyltransferase (ChAT) was detected with Burt and Silver methods, the automatic image analysissystem for biomedical application was applied in the quantitative analysisof AChE fiber and ChAT activity. MAIN OUTCOME MEASURES: ① Effect of Tiantai Ⅰ on the abilities of learning and memory in mice with Alzheimer disease; ② AChE fiberarea density in temporal cortex and hippocampal CA1 region; ③ ChAT ac tivity in Meynert nuclei of basal forebrain. RESULTS: ① Tiantai Ⅰ in ameliorating the abilities of learning and mem ory in mice with spontaneous Alzheimer disease: The error times in theTiantai Ⅰ 6.80 and 20.41 g/kg groups, normal control group and westerndrug control group were obviously fewer than that in the blank controlgroup (5.08±2.18, 3.77±1.48, 3.25±1.27, 6.17±3.12, 9.31±3.65, P ＜ 0.05). ② AChE fiber area density: The AChE fiber area densities in temporalcortex were obviously higher in the Tiantai Ⅰ 6.80 and 20.41 g/kg groups, normal control group and western drug control group than in the blankcontrol group (20.19±4.18, 25.01±6.44, 26.38±6.52, 19.44±4.33, 14.36±3.21, P ＜ 0.05). The AChE fiber area densities in hippocampal CA1 region were obviously higher in the Tiantai Ⅰ 20.41 g/kg groups and nor mal control group than in the blank control group, western drug controlgroup and Tiantai Ⅰ 6.80 g/kg group (13.88±3.07, 14.11±4.36, 6.57±2.30, 10.13±2.94, 9.63±2.40, P ＜ 0.05). ③ ChAT activity: It was obviously high er in the TiantaiI 20.41 and 6.80 g/kg groups, western drug control groupand normal control group than in the blank control group (0.683±0.179, 0.589±0.179, 0.552±0.173, 0.705±0.141, 0.354±0.131, P ＜ 0.05). CONCLUSION: Tiantai Ⅰ can significantly improve the disturbances oflearning and memory in mice with spontaneous Alzheimer disease, and itcan increase the activity of central cholinergic system in a dose-dependentmanner.
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BACKGROUND: The theory of free radical on onset of Alzheimer disease(AD) and the relationship between oxidative damage and onset of AD have made anti-oxidative drugs become the hot topic in researches. Whether does the action of natural Chinese herbs on anti-injury of oxygenic free radical produce intervention on AD?OBJECTIVE: To observe the effects of developed and researched tiantai I,the natural Chinese herb, on learning and memory function and metabolism of oxygenic free radical in the model of spontaneous AD.DESIGN: Completely randomized experimental study based on the experimental animals.SETTING: Clinical Institute of Chinese-Western integration of City-grade.MATERIALS: The experiment was performed in 2nd grade Animal Experimental Room of Clinical Institute of Chinese-Western integration in Shenzhen. Kunming male mice of clean grade were employed, at the age of 13 months, mass weighted varied from 40 to 50 g and from Experimental Animal Center of Zhongshan Medical University(Qualified No. 95A07).INTERVENTIONS: When the experimental animals were bred till 21 months, AD mice were screened from the aged animals according to International Universal Standards and Methods. The experimental animals were randomized into blank control, western-drug control, the small dosage of tiantai I group(small dosage group), the large dosage of tiantai I group(large dosage group) and learning and senile normal-memory group(senile normal group) in addition. In western-drug control, hydergine 0.6 mg/kg was administrated, and in small and large groups, tiantai I of 6. 80 g/kg and 20.41 g/kg were administrated respectively, continuously for 60 days. The double-distilled water of equal dosage was infused in both blank control and senile normal group.MAIN OUTCOME MEASURES: Step-down test was applied to determine learning and memory scores. Froze sections of brain tissue were prepared and immuno-histological-chemical method was used to display the superoxidative dismutase(SOD) activity in cerebral cortex and hippocampus. Ultrathin section of brain tissue in hippocampal CA1 was prepared and transmission electron microscope(TEM) was used to observe cellular lipofuscin in neuron. Automatic image analysis system for biomedical application was used to determine in quantity SOD and lipofuscin.RESULTS: Learning and memory of AD mice were significantly decreased ( P ＜ 0.01 ) . Tiantai I improved significantly learning and memory disturbance of mice in the model of spontaneous AD( P ＜ 0.05- 0.01),markedly enhanced SOD activity in central cerebral cortex and hippocampus ( P ＜ 0.05 - 0.01 ) and reduced cellular lipofuscin content of neuron( P＜ 0.05 -0.01 ). In addition, it provided certain of quantitative reaction.CONCLUSION: For the mice with spontaneous senile dementia, learning and memory as well as SOD activity of neuron in relevant cerebral areas are decreased remarkably and lipofuscin is increased significantly. Tiantai I acts significantly in pharmacodynamics on anti-injury of oxygenic free radical,increases significantly SOD activity, decreases lipofuscin content and improves the ultrastructure of neuron in cortex and hippocampus in AD mice.
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BACKGROUND:Neuron nitrogen monoxide(NO) is related to synaptic plasticity and is the key transmitter of normal learning and memory.Neuronal nitric oxide synthase(nNOS) is the biological synthetic enzyme of NO in neurons.The reports have been fewer yet on cerebral nNOS activity in Alzheimer disease(AD) patients,especially on changes in positive neural fibers of nNOS in temporal lobe cortex and hippocapmus CA1 region. OBJECTIVE:To observe cerebral nNOS activity in AD patients, especially on changes in positive neural fibers of nNOS in temporal lobe cortex and hippocapmus CA1 region and effects of Tiantai Ⅰ on nNOS activity. DESIGN:A randomized and controlled trial. SETTING:Institute of Collaboration of Chinese and Western Medicine, Shenzhen City. MATERIALS:The experiment was performed in No.2 Grade Animal Experimental Room of Institute of Collaboration of Chinese and Western Medicine,Shenzhen City.A total of 65 Kunming mice were employed in the experiment. METHODS:The experimental animals were bred till 21 months old and the mice with senile dementia were screened from the aged ones according to the international general standards and methods.The experimental animals were randomized into 4 groups,named as senile dementia group,western drug control,group with small dosage of Tiantai Ⅰ and group with large dosage of Tiantai Ⅰ .In addition,a group with normal learning and memory of the senile mice(simply called senile normal group) was designed.There were 13 mice in each group.In western drug control,hydergine 0.6 mg/kg was prescribed; in the groups of small and large dosage of Tiantai Ⅰ ,the dosages were 6.80 g/kg and 20.41 g/kg respectively,continuously for 60 days.In senile normal group and senile dementia group,the bi distilled water of equal dosage was applied for perfusion.The scores of learning and memory were determined by step down test. Slices of brain tissues were prepared with frozen,nNOS activity was displayed by NBT histochemistry method and the analysis was done systematically with auto micrography in quantity. RESULTS:Learning and memory were significantly decreased in mice with senile dementia(P< 0.01),the positive reaction of nNOS was remarkably weaker compared with normal aged mice.It was the first time to discover that the surface densities of nNOS positive neural fibers in temporal lobe cortex and hippocampus CA1 region [(9.48± 2.09),(5.11± 2.74) respectively]were remarkably lower than those of the normal aged mice [(17.75± 3.47),(11.95± 2.00) respectively](P< 0.01). Simultaneously, it was also the first time to discover that normal correlation was apparent between nNOS activity of cortex and hippocampus and the scores of learning and memory.Tiantai Ⅰ could significantly increase the surface densities of nNOS positive neural fibers in cortex and hippocampus in senile dementia mice, of which, the values of the small dosage were (11.57± 2.44) and (8.18± 2.92) respectively and of the large dosage were (12.59± 2.88) and (9.33± 2.18) respectively,(P< 0.01- 0.05). CONCLUSION:The hypo activity of central nNOS in senile dementia mice resulted in significantly decreased positive neural fibers of nNOS in cerebral cortex and hippocampus.The surface densities of nNOS positive neural fibers in temporal lobe cortex and hippocampus CA1 region were significantly related to the scores of learning and memory.Tiantai Ⅰ could remarkably improve the disturbance of learning and memory in spontaneous senile dementia model and enhance its nNOS activity in cerebral cortex and hippocampus.
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BACKGROUND: The synaptic plasticity of central nerves is the key basis of behavior-dependent learning and memory, there have been few reports about weather the Chinese herbal medicine for treatment of Alzheimer disease is through enhancing synaptic plasticity to improve learning and memory.OBJECTIVE: To investigate the effect of tiantai No. 1 recipe on learning and memory function and synaptic plasticity of spontaneous Alzheimer disease models.DESIGN: A randomized and controlled trial.SETTING: Institute of Combination of Traditional Chinese Medicine and Western Medicine of Shenzhen.MATERIALS: The experiment was completed at the grade Ⅱ Laboratory of Institute of Combination of Traditional Chinese Medicine and Western Medicine of Shenzhen City. The experimental animals were mice of Kunming strain.METHODS: Totally 52 Kunming mice, aged 21 months, with spontaneous Alzheimer disease (memory impairment) were chosen, and at random divided into 4 groups: senile dementia, western drug control, tiantai No. 1recipe 6.80 g/kg and 20.41 g/kg groups; besides 13 mice with normal learning and memory power were chosen as normal senile group. Hydergine 0.6 mg/kg was given for mice in western drug control group, tiantai No. 1recipe 6.80 g/kg and 20.41 g/kg were respectively given for mice in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups. The medicines above were all prepared into 0.5 mL solution and given by gavage, for consecutive 60 days. The mice in normal senile and senile dementia groups were given double distilled water of the same volume. The learning memory power was tested by jumping stand; ultrathin cerebral section of hippocampal CA1 region was examined with transmission electron microscope,and some parameters of synaptic plasticity was also quantitatively detected with automatic micrographic analysis system.MAIN OUTCOME MEASURES: ① The effect of tiantai No. 1 recipe in different doses on learning and memory of spontaneous Alzheimer disease mice. ② Observation of synapse with electron microscope and quantitative examination stereologically.RESULTS: All animals involved in the trial entered the final result analysis without any loss. ① The effect of tiantai No. 1 recipe in different doses on learning and memory of spontaneous Alzheimer disease mice: The error numbers of learning and memory in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups were smaller than that in senile dementia group,and the error number of learning in tiantai No. 1 recipe 6.80 g/kg group was smaller than that in tiantai No. 1 recipe 20.41 g/kg group. The escape latent periods of learning training was smaller than that in senile dementia group and the safe platform latent periods of memory in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups were longer than that in senile dementia group, and the safe platform latent period of memory in tiantai No. 1 recipe 6.80 g/kg group was smaller than that in 20.41 g/kg group. ② Observation of synapse with electron microscope and quantitative examination stereologically: Compared with the mice in normal senile group, for mice in senile dementia group, there occurred degenerative phenomena: the number of synapses reduced, a part of synaptic clefts were not clear, the synaptic junction was interrupted, and he sizes of synaptic vesicles were different; in the rest groups there were also degenerative phenomena, however the degrees were milder than those in the model group. Tiantai No. 1 recipe could remarkably raise the number density and sectional density of synapses in the hippocampal CA1 region, and the increased extents in 20.41 g/kg tiantai No.1 recipe was larger than those in 6.80 g/kg tiantai No. 1 recipe.CONCLUSION: Tiantai No. 1 recipe can remarkably improve their learning and memory impairments, which may be related with that it promotes re-generation of synapse and improves the plasticity of cerebral synapses;and the action presented a relationship of dose-effect.
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BACKGROUND: The early and medium medication has been proved to have a certain effect to patients with Alzheimer disease, can delay its development.OBJECTIVE: To investigate the effect of No. 1 recipe on learning and memory function of spontaneous Alzheimer disease models.DESIGN: A randomized and controlled trial.SETTING: Shenzhen City Institute of Integration of Traditional Chinese and Western Medicine.MATERIALS: The experiment was completed at the grade Ⅱ Laboratory of Shenzhen City Institute of Integration of Traditional Chinese and Western Medicine. The experimental animals were mice of Kunming strain.METHODS: Totally 52 Kunming mice, aged 21 months, with spontaneous Alzheimer disease (memory impairment) were chosen, and at random divided into 4 groups: senile dementia, western drug control, tiantai No. 1recipe 6.80 g/kg and 20.41 g/kg groups; besides 13 mice with normal learning and memory power were chosen as normal senile group. Hydergine 0.6 mg/kg was given for mice in western drug control group, tiantai No. 1recipe (orthogonally optimized) 6.80 g/kg and 20.41 g/kg were respectively given for mice in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups.The medicines above were all prepared into 0.5 mL solution and given by garage, for consecutive 60 days. The mice in normal senile and senile dementia groups were given double distilled water of the same volume. The learning memory power was tested by jumping stand, comparing the numbers of learning error and memory error, latent escaping periods of learning training and latent safe platform of memory test.MAIN OUTCOME MEASURES: The improving effect of tiantai No. 1recipe on learning and memory function of spontaneous Alzheimer disease mice.RESULTS: All animals involved in the trial entered the final result analysis without any loss. The error numbers of learning and memory in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups were smaller than that in senile dementia group (5.08±2.18, 3.77±1.48, 9.31±3.65; 1.85±1.21, 1.54±0.88,4.27±2.58), and the error number of learning in tiantai No. 1 recipe 6.80 g/kg group was more than that in tiantai No. 1 recipe 20.41 g/kg group. The escape latent period of learning training was smaller than that in the senile dementia group and the safe platform latent period of memory in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups were longer than that in senile dementia group [(18.9±7.5), (19.9±5.9), (33.7±9.3); (91.7±32.0),(101.5±40.9), (43.6±20.7)s], and the safe platform latent period of memory in tiantai No. 1 recipe 6.80 g/kg group was smaller than that in 20.41 g/kg group.CONCLUSION: There was obvious intellectual disturbance in spontaneous Alzheimer disease mice, tiantai No. 1 recipe can remarkably improve their learning and memory impairments, with a relationship of dose-effect.