1.Effect of Invigorating Spleen,Eliminating Stasis,Heat-Clearing Drugs on the Quality of Ulcer Healing and Expression of EGFR mRNA in the Rat Model of Gastric Ulcer
Yongfeng ZHANG ; Zhengzhi WU ; Xiuqin JIA
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(12):-
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.
2.Effects of Tiantai Ⅰ on the activity of central cholinergic system in mice with spontaneous Alzheimer disease
Zhengzhi WU ; Ming LI ; Yaofang LI ; Xiuqin JIA ; Yongfeng ZHANG
Chinese Journal of Tissue Engineering Research 2006;10(35):163-165
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.
3.Neural Mechanisms of Forward Conditional Reasoning:A Functional MRI Study
Peipeng LIANG ; Xiuqin JIA ; Lin SHI ; Defeng WANG ; Kuncheng LI
Chinese Journal of Medical Imaging 2015;(4):241-244
Purpose Two kinds of forward conditional reasoning including modus ponens (MP) and denial of the antecedent (DA) are the most common deductive reasoning in daily life. The present study aimed to explore the difference between the two neural machanisms. Materials and Methods Three kinds of tasks including MP, DA, and baseline were administrated to 14 undergraduates [7 males and 7 females, mean age (23.4±1.3) years] by using functional magnetic resonance imaging (fMRI) technique. fMRI data was analyzed. Results The reaction times were significantly slower for MP and DA than that of BS (P<0.001), while the effect was not significant for accuracy. In contrast to baseline, both MP and DA tasks activated the left inferior frontal cortex (BA 9), inferior parietal lobule (BA 40), and postcentral gyrus. Bilateral caudate significant activation was found in MP task when compared with DA, whereas the frontal, parietal, occipital lobe and anterior cingulate cortex were acivated more in the DA than in MP task. Conclusion Both forward conditional reasoning of MP and DA commonly activate the left frontoparietal cortex, while significant dissociations can be seen in the regions of caudate and bilateral frontal, parietal and occipital. These results firstly demonstrates the different neural mechanism underlying the two forward conditional reasoning, which might help to further uncover the brain mechanism of conditional reasoning.
4.Influence of tiantai No. 1 recipe on learning and memory function of spontaneous Alzheimer disease models
Zhengzhi WU ; Ming LI ; Xiuqin JIA ; Yongfeng ZHANG
Chinese Journal of Tissue Engineering Research 2005;9(45):180-181
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.
5.Changes of neuronal nitric oxide synthase in relevant cerebral regions in spontaneous senile dementia model and regulation of Tiantai Ⅰ
Zhengzhi WU ; Ming LI ; Xiuqin JIA ; Yongfeng ZHANG
Chinese Journal of Tissue Engineering Research 2005;9(4):244-247
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.
6.Effect of nerve growth factor on adrenergic nerve in rals after myocardial infarction
Xiuqin NI ; Xing LI ; Jia FENG ; Linghui HAO ; Changwei JI
Clinical Medicine of China 2008;24(10):991-993
Objective To confirmthe protective effect of nerve growth factor (NGF)on cadiac adrenergie nerve in rats after acute myocardial infarction(AMI)and its mechanism.Methods 120 Wister rats were divided in-to sham-operated group.AMI group and NGF group.The samples were taken after6 h, d,4 d-7 d and 14 d sepa-lately.Immunohistochemistry method WaS used to show the distribution of adrenergic nerve fibers.The density of them were calculated by Medical Pathobgical Image Analysis Processing System.Results ①The densities of adrenergic nerve fibers in AMI group were obviously lower than that of sham-operated group greatly in 6 h and 2 d.4 d-7 d and14 d later.②The densities of adrenergic innervation in NGF group were obviously higher than that of AMI group 7 dand 14 d later.Conclusion NGF has protection effect 011 cardLac adrenergic nerve after AMI in the rat.
7.Correlation study of methylenetetrahydrofotate reductase C677T polymorphisms and H-type hypertension and increased homocysteine levels
Jia WANG ; Shuling WANG ; Dan ZENG ; Na ZHANG ; Xiuqin HONG
Chinese Journal of Cerebrovascular Diseases 2015;(11):581-586
Objective To investigate the relationship between methylenetetrahydrofotate reductase (MTHFR)C677T polymorphisms and H-type hypertension and increased plasma homocysteine (Hcy) levels. Methods From September 2013 to June 2014,4 012 permanent residents aged ≤30 year from 12 natural villages or communities in 6 regions of Hunan province were extracted according to the cluster random sampling method. Using computer random number table,571 residents were randomly selected as the research objects. According to the blood pressure and Hcy levels,571 residents were divided into 3 groups:a common hypertension group (n = 190),an H-type hypertension group (n = 94),and a normal blood pressure group (n = 287 ). Amplification refractory mutation system-polymerase chain reaction (ARMS-PCR)method was used to detect the MTHFR C677T polymorphisms in all the research objects and the penotyping was performed. Hcy levels were detected at the same time. Results There were significant differences in recessive model (CC + CT,TT)genotype frequencies among the H-type hypertension group (n = 66[70. 2%],n = 28[29. 8%]),common hypertension group (n = 156[82. 1%],n = 34[17. 9%]), and normal blood pressure group (n = 235[81. 9%],n = 52[18. 1%])(χ2 = 6. 797,P = 0. 033),and there were no significant differences in CC,CT,and TT genotype frequencies among the 3 groups (P >0. 05). In the recessive model,there were significant differences in TT genotype frequencies between the H-type hypertension group and the normal blood pressure group or the common hypertension group (χ2 = 5. 812,P = 0. 016;χ2 = 5. 212,P = 0. 022). There was no significant difference in TT genotype frequencies between the common hypertension group and the normal blood pressure group (P > 0. 05). The CC + CT and TT genotype Hcy levels of the MTHFR C677T recessive model in the H-type hypertension group were 17. 1 ±1. 6 and 19. 0 ±2. 9 μmol/ L respectively. There was significant difference between the genotypes (t = - 3. 115,P = 0. 004). The logistic regression analysis of MTHFR C677T recessive model genotype showed that after adjusting for sex and age,the residents with recessive model TT genotype had higher risk of H-type hypertension (OR,1. 946,95% CI 1. 172 -3. 232,P = 0. 01). Conclusion The TT MTHFR C677T gene mutation in this population may be an important genetic factor for the increased Hcy levels and the onset of H-type hypertension.
8.Influence of tiantai No. 1 recipe on synaptic plasticity of the hippocampal CA1 region of spontaneous Alzheimer disease models: Quantitative research of ultrastructure
Ming LI ; Zhengzhi WU ; Yaofang LI ; Yongfeng ZHANG ; Xiuqin JIA ; Manyin CHEN
Chinese Journal of Tissue Engineering Research 2005;9(45):182-184
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.
9.Quantitative study of tiantai I on superoxidative dismutase and lipofuscin in relevant cerebral areas of spontaneous Alzheimer disease in mice
Zhengzhi WU ; Ming LI ; Yaofang LI ; Yongfeng ZHANG ; Xiuqin JIA ; Manyin CHEN
Chinese Journal of Tissue Engineering Research 2005;9(13):178-181
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.
10.In vitro isolation of intestinal side population cells from newborn mice
Yongfeng ZHANG ; Jin YANG ; Zhengzhi WU ; Xiuqin JIA ; Ming LI ; Yinghong LI
Chinese Journal of Tissue Engineering Research 2010;14(27):5115-5118
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.