Aim： To access beta-endorphin levels in serum as well as seminal plasma in different infertile male groups. Methods： Beta-endorphin was estimated in the serum and seminal plasma by enzyme-linked immunosorbent assay ...Aim： To access beta-endorphin levels in serum as well as seminal plasma in different infertile male groups. Methods： Beta-endorphin was estimated in the serum and seminal plasma by enzyme-linked immunosorbent assay （ELISA） method in 80 infertile men equally divided into four groups： non-obstructive azoospermia （NOA）, obstructive azoospermia （OA）, congenital bilateral absent vas deferens （CBVAD） and asthenozoospermia. The results were compared to those of 20 normozoospermic proven fertile men. Results： There was a decrease in the mean levels of betaendorphin in the seminal plasma of all successive infertile groups （mean ± SD： NOA 51.30 ± 27.37, OA 51.88 ± 9.47, CBAVD 20.36 ± 13.39, asthenozoospermia 49.26 ± 12.49 pg/mL, respectively） compared to the normozoospermic fertile control （87.23 ± 29.55 pg/mL）. This relation was not present in mean serum level of beta-endorphin between four infertile groups （51.09 ± 14.71, 49.76 ± 12.4, 33.96 ± 7.2, 69.1 ± 16.57 pg/mL, respectively） and the fertile control group （49.26 ± 31.32 pg/mL）. The CBVAD group showed the lowest seminal plasma mean level of beta-endorphin. Testicular contribution of seminal beta-endorphin was estimated to be approximately 40%. Seminal beta-endorphin showed significant correlation with the sperm concentration （r = 0.699, P = 0.0188） and nonsignificant correlation with its serum level （r = 0.375, P = 0.185） or with the sperm motility percentage （r = 0.470, P = 0.899）. Conclusion： The estimation of beta-endorphin alone is not conclusive to evaluate male reproduction as there are many other opiates acting at the hypothalamic pituitary gonadal axis.展开更多
β-endorphin is the most actively endogenous substance of cerebral endorphin. When combined with opiate receptor specially, it manifests a strong morphine-like activity and can decrease sensitivity of central nervous ...β-endorphin is the most actively endogenous substance of cerebral endorphin. When combined with opiate receptor specially, it manifests a strong morphine-like activity and can decrease sensitivity of central nervous system to carbon dioxide so as to inhibit breath. OBJECTIVE： To observe the changes of content of plasma β-endorphin in neonates with severe asphyxia after naloxone treatment in a large dosage. DESIGN： Randomized controlled observation. SETTINGS： Department of Pediatrics, Shenzhen Shajing People＇s Hospital; Center of Pediatrics, Guangzhou Zhujiang Hospital. PARTICIPANTS： A total of 97 neonates with severe asphyxia including 57 boys and 40 girls were selected from Neonatal Intensive Care Unit, Department of Pediatrics, Shenzhen Shajing People＇s Hospital from January 2004 to November 2005. Their gestational age was （38±3） weeks, body mass was （3.2±1.7） kg, and hospitalization duration was （2.8±2.3） hours. All neonates met the diagnostic criteria of with severe asphyxia and all their parents provided the confirmed consent. METHODS： All neonates were treated with inspired oxygen, sedation, stopping terror, decreasing cranial pressure, maintaining a well blood perfusion and normal level of blood glucose （about 5.0 retool/L）. After hospitalization, 0.1 mg/（kg·d） naloxone hydrochloride （Beijing Sihuan Pharmaceutical Technology Co., Ltd.; certification： HI0900021; bullet preparation; 0.4 mg/ampoule） was intravenously dribbled into neonates for 4 - 6 hours, 14 days in total. 2 mL blood was collected from radial artery in neonates at the beginning of hospitalization and at 3 days after naloxone treatment, put in aprotinin-pre-cool tube, mixed evenly, and centrifuged at hypothermia. Plasma was maintained in refrigerator at - 70 ℃. The kit was provided by Neurobiology Department of Shanghai Second Military Medical University of Chinese PLA. Concentration of plasma β-endorphin was measured by using radio-immunity assay.All data were expressed as Mean ± SD and results were compared with paired t test. MAIN OUTCOME MEASURE： Concentration of plasma β-endorphin. RESULTS： All 97 neonates were involved in the final analysis. Concentration of plasma β-endorphin in neonates with severe asphyxia was lower after treatment as compared with that before treatment, and there was significant difference （t = 10.31, P 〈 0.01 ）. CONCLUSION： Naloxone can decrease level of plasma β-endorphin in neonates with severe asphyxia.展开更多
BACKGROUND： Several studies have confirmed that endothelin and endorphin are involved in the occurrence of cerebral vasospasm. However, the correlation of these factors to acute cerebral infarction-related risk facto...BACKGROUND： Several studies have confirmed that endothelin and endorphin are involved in the occurrence of cerebral vasospasm. However, the correlation of these factors to acute cerebral infarction-related risk factors needs to be confirmed. OBJECTIVE： To detect endothelin-1 （ET-1） and beta-endorphin （β -EP） levels in plasma of patients with acute cerebral infarction, and to analyze the correlations of these factors to smoking, alcohol abuse, hypertension, diabetes mellitus, diseased region, diseased degree, gender, and other factors related to acute cerebral infarction. DESIGN： A case-control observation. SETTING： First Department of Neurology, Guangdong Hospital of Traditional Chinese Medicine; Department of Neurology, Second Affiliated Hospital of Sun Yat-sen University. PARTICIPANTS： Sixty-nine inpatients with acute cerebral infarction were admitted to the Department of Neurology, Second Affiliated Hospital of Sun Yat-sen University （March 2003-January 2004） and First Department of Neurology, Guangdong Hospital of Traditional Chinese Medicine （March July 2004） and recruited for this study. All 69 inpatients corresponded to the diagnosis criteria of acute cerebral infarction, formulated in the National Working Conference of Cerebrovascular Disease in 1998, and were confirmed as acute cerebral infarction by CT/MRI. The patient group consisted of 35 males [（644- 12） years old] and 34 females[ （674- 13 ） years old]. Among them, 9 patients were smokers, 7 were alcohol users, 48 had a history of hypertension, and 16 had a history of diabetes mellitus. CT/MRI examinations revealed that 35 patients presented with left focus sites, 11 with right ones and 23 with bilateral ones. Following attack, 24 patients had Barthel Index Scale grading 〈 40 points, 21 patients 40-50 points, and 24 patients 〉 60 points. An additional 59 healthy individuals, who received health examinations simultaneously, were included as controls. Among the control subjects, there were 37 males [（62±10） years old] and 22 females [（65±11） years old]. Among them, 7 patients were smokers, and 6 were alcohol users. All controls had no history of stroke, hypertension, or diabetes mellitus. Informed consents of laboratory measurements were obtained from all subjects, and this study was approved by the Hospital Ethics Committee. METHODS： ① Following admission, all subjects were scored by Barthel Index Scale （BIS） and Hamilton Depression Scale. Meanwhile, hypertension, diabetes mellitus, gender, smoking, drinking, and other conditions were recorded. CT/MRI examination was conducted to identify the focus site.②On the 2^nd day after admission, ET-1 and β -EP plasma levels were measured with an automatic ET-1 and β -EP analysis kit. MAIN OUTCOME MEASURES： ET-1 and β -EP plasma levels and their correlation to acute cerebral infarction-related factors. RESULTS： Sixty-nine patients with acute cerebral infarction, and an additional 59 healthy individuals participated in the final analysis. β ET-1 [（63.80±27.65） ng/L vs. （46.50±9.36） ng/L, P 〈 0.05] and β - EP [（94.18±33.94） mg/L vs. （51.87±23.43） mg/L, P 〈 0.05] levels of the patient group were obviously higher than respective values of the control group. ② The ET-1 and β -EP levels of patients with cerebral infarction did not correlate to hypertension, diabetes mellitus, BIS, depression, cerebral infarct focus, disease course, gender, smoking or drinking （P 〉 0.05）. CONCLUSION： The ET-I and β-EP levels of patients with acute cerebral infarction increased, but they were not obviously associated with disease course, blood pressure, blood glucose, BIS, or other common cerebral infarction-related factors.展开更多
许多实证研究表明,单个股票的Beta和其收益之间并没有显著的相关关系,由此否认传统GAPM模型所揭示的风险-收益关系。但在所用模型中的预期收益和实证检验中运用的已实现收益是两个不同的概念,本文运用Pettengill et al提出的条件CAPM模...许多实证研究表明,单个股票的Beta和其收益之间并没有显著的相关关系,由此否认传统GAPM模型所揭示的风险-收益关系。但在所用模型中的预期收益和实证检验中运用的已实现收益是两个不同的概念,本文运用Pettengill et al提出的条件CAPM模型对上海股市重新作检验,研究发现在牛市中Beta和收益呈现出显著的正相关关系,在熊市中Beta和收益呈现出显著的负相关关系,且两者显示出一定的对称性。研究结果表明,在上海股市中条件CAPM模型具有相当的应用空间。展开更多
Mounting evidence indicates that amyloid β protein（Aβ） exerts neurotoxicity by disrupting the blood-brain barrier（BBB） in Alzheimer＇s disease. Hyperoside has neuroprotective effects both in vitro and in vivo ag...Mounting evidence indicates that amyloid β protein（Aβ） exerts neurotoxicity by disrupting the blood-brain barrier（BBB） in Alzheimer＇s disease. Hyperoside has neuroprotective effects both in vitro and in vivo against Aβ. Our previous study found that hyperoside suppressed Aβ1-42-induced leakage of the BBB, however, the mechanism remains unclear. In this study, bEnd.3 cells were pretreated with 50, 200, or 500 μM hyperoside for 2 hours, and then exposed to Aβ1-42 for 24 hours. Cell viability was determined using 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide assay. Flow cytometry and terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling assay were used to analyze cell apoptosis. Western blot assay was carried out to analyze expression levels of Bax, Bcl-2, cytochrome c, caspase-3, caspse-8, caspase-9, caspase-12, occludin, claudin-5, zonula occludens-1, matrix metalloproteinase-2（MMP-2）, and MMP-9. Exposure to Aβ1-42 alone remarkably induced bEnd.3 cell apoptosis; increased ratios of cleaved caspase-9/caspase-9, Bax/Bcl-2, cleav ed caspase-8/caspase-8, and cleaved caspase-12/caspase-12; increased expression of cytochrome c and activity of caspase-3; diminished levels of zonula occludens-1, claudin-5, and occludin; and increased levels of MMP-2 and MMP-9. However, hyperoside pretreatment reversed these changes in a dose-dependent manner. Our findings confirm that hyperoside alleviates fibrillar Aβ1-42-induced BBB disruption, thus offering a feasible therapeutic application in Alzheimer＇s disease.展开更多
BACKGROUND： Neuronal loss, synapse mutilation, and increasing malnourished axons are pathologically related to Alzheimer＇s disease. Microtubule-associated protein 2 （MAP2） is of importance for neuronal, axonal, an...BACKGROUND： Neuronal loss, synapse mutilation, and increasing malnourished axons are pathologically related to Alzheimer＇s disease. Microtubule-associated protein 2 （MAP2） is of importance for neuronal, axonal, and dendritic generation, extension, and stabilization, as well as for the regulation of synaptic plasticity. OBJECTIVE： To investigate the antagonistic effects of natural-cerebrolysin-containing serum on beta amyloid protein 1-40 （Aβ1-40）-induced neurotoxicity from the standpoints of cell proliferation, synaptogenesis, and cytoskeleton formation （MAP2 expression）. DESIGN, TIME AND SETTING： A paralleled, controlled, neural cell, and molecular biology experiment was performed at the Institute of Integrated Chinese and Western Medicine, Shenzhen Hospital, Southern Medical University between February 2006 and April 2008. MATERIALS： PC12 cells, derived from the rat central nervous system, were purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China. A β1-40 was provided by Sigma, USA. Natural-cerebrolysin was provided by Shenzhen Institute of Integrated Chinese and Western Medicine, China. The natural-cerebrolysin was predominantly composed of Renshen （Radix Ginseng）, Tianma （Rhizoma Gastrodiae）, and Yixingye （Ginkgo Leaf） in a proportion of 1：2：2. Following conventional water extraction technology, an extract （1：20） was prepared. Each gram of extract equaled 20 grams of crude drug. In a total of 12 adult male New Zealand rabbits, six underwent intragastric administration of natural-cerebrolysin extract for 1 month to prepare natural-cerebrolysin-containing serum, and the remaining six rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： An AIzheimer＇s disease in vitro model was induced in PC12 cells using Aβ1-40. The cells were incubated with varying doses of natural-cerebrolysin-containing serum （2.5%, 5%, and 10%）. Normal blank serum-treated PC12 cells served as a blank control group. MAIN OUTCOME MEASURES： Through the use of inverted phase contrast microscope, cell morphology and neurite growth were observed, neurite length was measured, and the percentage of neurite-positive cells was calculated. Cell proliferation rate was determined by MTT assay, and MAP 2 expression was detected by fluorescent immunocytochemistry. RESULTS： Following Aβ1-40 treatments, some PC12 cells were apoptotic/dying, and only a few short neurites were observed. Following interventions with natural-cerebrolysin-containing serum, the PC12 cells proliferated, there was an increased number of neurites, and neurite length was enhanced. After middle- and high-dose natural-cerebrolysin treatments, the percentage of neurite-positive cells, as well as the average length of neurites, was significantly greater than the normal blank serum-treated PC12 cells （P 〈 0.05 or P 〈 0.01）. Compared with the blank control group, MAP2 expression in the Aβ1-40-treated PC12 cells was significantly inhibited, and the cell proliferation rate was significantly decreased （P 〈 0.01）. Following incubations with natural-cerebrolysin-containing serum, MAP2 expression and cell proliferation rate in the PC12 cells were significantly increased in a dose-dependent manner, compared with treatments with blank control serum （P 〈 0.05 or P 〈 0.01 ）. CONCLUSION： Natural-cerebrolysin exhibited antagonistic effects on neurotoxicity in Aβ1-40 induced Alzheimer＇s disease in vitro models. These effects were likely related to cell proliferation and the upregulation of intracellular MAP2 expression.展开更多
文摘Aim： To access beta-endorphin levels in serum as well as seminal plasma in different infertile male groups. Methods： Beta-endorphin was estimated in the serum and seminal plasma by enzyme-linked immunosorbent assay （ELISA） method in 80 infertile men equally divided into four groups： non-obstructive azoospermia （NOA）, obstructive azoospermia （OA）, congenital bilateral absent vas deferens （CBVAD） and asthenozoospermia. The results were compared to those of 20 normozoospermic proven fertile men. Results： There was a decrease in the mean levels of betaendorphin in the seminal plasma of all successive infertile groups （mean ± SD： NOA 51.30 ± 27.37, OA 51.88 ± 9.47, CBAVD 20.36 ± 13.39, asthenozoospermia 49.26 ± 12.49 pg/mL, respectively） compared to the normozoospermic fertile control （87.23 ± 29.55 pg/mL）. This relation was not present in mean serum level of beta-endorphin between four infertile groups （51.09 ± 14.71, 49.76 ± 12.4, 33.96 ± 7.2, 69.1 ± 16.57 pg/mL, respectively） and the fertile control group （49.26 ± 31.32 pg/mL）. The CBVAD group showed the lowest seminal plasma mean level of beta-endorphin. Testicular contribution of seminal beta-endorphin was estimated to be approximately 40%. Seminal beta-endorphin showed significant correlation with the sperm concentration （r = 0.699, P = 0.0188） and nonsignificant correlation with its serum level （r = 0.375, P = 0.185） or with the sperm motility percentage （r = 0.470, P = 0.899）. Conclusion： The estimation of beta-endorphin alone is not conclusive to evaluate male reproduction as there are many other opiates acting at the hypothalamic pituitary gonadal axis.
文摘β-endorphin is the most actively endogenous substance of cerebral endorphin. When combined with opiate receptor specially, it manifests a strong morphine-like activity and can decrease sensitivity of central nervous system to carbon dioxide so as to inhibit breath. OBJECTIVE： To observe the changes of content of plasma β-endorphin in neonates with severe asphyxia after naloxone treatment in a large dosage. DESIGN： Randomized controlled observation. SETTINGS： Department of Pediatrics, Shenzhen Shajing People＇s Hospital; Center of Pediatrics, Guangzhou Zhujiang Hospital. PARTICIPANTS： A total of 97 neonates with severe asphyxia including 57 boys and 40 girls were selected from Neonatal Intensive Care Unit, Department of Pediatrics, Shenzhen Shajing People＇s Hospital from January 2004 to November 2005. Their gestational age was （38±3） weeks, body mass was （3.2±1.7） kg, and hospitalization duration was （2.8±2.3） hours. All neonates met the diagnostic criteria of with severe asphyxia and all their parents provided the confirmed consent. METHODS： All neonates were treated with inspired oxygen, sedation, stopping terror, decreasing cranial pressure, maintaining a well blood perfusion and normal level of blood glucose （about 5.0 retool/L）. After hospitalization, 0.1 mg/（kg·d） naloxone hydrochloride （Beijing Sihuan Pharmaceutical Technology Co., Ltd.; certification： HI0900021; bullet preparation; 0.4 mg/ampoule） was intravenously dribbled into neonates for 4 - 6 hours, 14 days in total. 2 mL blood was collected from radial artery in neonates at the beginning of hospitalization and at 3 days after naloxone treatment, put in aprotinin-pre-cool tube, mixed evenly, and centrifuged at hypothermia. Plasma was maintained in refrigerator at - 70 ℃. The kit was provided by Neurobiology Department of Shanghai Second Military Medical University of Chinese PLA. Concentration of plasma β-endorphin was measured by using radio-immunity assay.All data were expressed as Mean ± SD and results were compared with paired t test. MAIN OUTCOME MEASURE： Concentration of plasma β-endorphin. RESULTS： All 97 neonates were involved in the final analysis. Concentration of plasma β-endorphin in neonates with severe asphyxia was lower after treatment as compared with that before treatment, and there was significant difference （t = 10.31, P 〈 0.01 ）. CONCLUSION： Naloxone can decrease level of plasma β-endorphin in neonates with severe asphyxia.
文摘BACKGROUND： Several studies have confirmed that endothelin and endorphin are involved in the occurrence of cerebral vasospasm. However, the correlation of these factors to acute cerebral infarction-related risk factors needs to be confirmed. OBJECTIVE： To detect endothelin-1 （ET-1） and beta-endorphin （β -EP） levels in plasma of patients with acute cerebral infarction, and to analyze the correlations of these factors to smoking, alcohol abuse, hypertension, diabetes mellitus, diseased region, diseased degree, gender, and other factors related to acute cerebral infarction. DESIGN： A case-control observation. SETTING： First Department of Neurology, Guangdong Hospital of Traditional Chinese Medicine; Department of Neurology, Second Affiliated Hospital of Sun Yat-sen University. PARTICIPANTS： Sixty-nine inpatients with acute cerebral infarction were admitted to the Department of Neurology, Second Affiliated Hospital of Sun Yat-sen University （March 2003-January 2004） and First Department of Neurology, Guangdong Hospital of Traditional Chinese Medicine （March July 2004） and recruited for this study. All 69 inpatients corresponded to the diagnosis criteria of acute cerebral infarction, formulated in the National Working Conference of Cerebrovascular Disease in 1998, and were confirmed as acute cerebral infarction by CT/MRI. The patient group consisted of 35 males [（644- 12） years old] and 34 females[ （674- 13 ） years old]. Among them, 9 patients were smokers, 7 were alcohol users, 48 had a history of hypertension, and 16 had a history of diabetes mellitus. CT/MRI examinations revealed that 35 patients presented with left focus sites, 11 with right ones and 23 with bilateral ones. Following attack, 24 patients had Barthel Index Scale grading 〈 40 points, 21 patients 40-50 points, and 24 patients 〉 60 points. An additional 59 healthy individuals, who received health examinations simultaneously, were included as controls. Among the control subjects, there were 37 males [（62±10） years old] and 22 females [（65±11） years old]. Among them, 7 patients were smokers, and 6 were alcohol users. All controls had no history of stroke, hypertension, or diabetes mellitus. Informed consents of laboratory measurements were obtained from all subjects, and this study was approved by the Hospital Ethics Committee. METHODS： ① Following admission, all subjects were scored by Barthel Index Scale （BIS） and Hamilton Depression Scale. Meanwhile, hypertension, diabetes mellitus, gender, smoking, drinking, and other conditions were recorded. CT/MRI examination was conducted to identify the focus site.②On the 2^nd day after admission, ET-1 and β -EP plasma levels were measured with an automatic ET-1 and β -EP analysis kit. MAIN OUTCOME MEASURES： ET-1 and β -EP plasma levels and their correlation to acute cerebral infarction-related factors. RESULTS： Sixty-nine patients with acute cerebral infarction, and an additional 59 healthy individuals participated in the final analysis. β ET-1 [（63.80±27.65） ng/L vs. （46.50±9.36） ng/L, P 〈 0.05] and β - EP [（94.18±33.94） mg/L vs. （51.87±23.43） mg/L, P 〈 0.05] levels of the patient group were obviously higher than respective values of the control group. ② The ET-1 and β -EP levels of patients with cerebral infarction did not correlate to hypertension, diabetes mellitus, BIS, depression, cerebral infarct focus, disease course, gender, smoking or drinking （P 〉 0.05）. CONCLUSION： The ET-I and β-EP levels of patients with acute cerebral infarction increased, but they were not obviously associated with disease course, blood pressure, blood glucose, BIS, or other common cerebral infarction-related factors.
文摘许多实证研究表明,单个股票的Beta和其收益之间并没有显著的相关关系,由此否认传统GAPM模型所揭示的风险-收益关系。但在所用模型中的预期收益和实证检验中运用的已实现收益是两个不同的概念,本文运用Pettengill et al提出的条件CAPM模型对上海股市重新作检验,研究发现在牛市中Beta和收益呈现出显著的正相关关系,在熊市中Beta和收益呈现出显著的负相关关系,且两者显示出一定的对称性。研究结果表明,在上海股市中条件CAPM模型具有相当的应用空间。
基金financially supported by the National Natural Science Foundation of China,No.81573771the Natural Science Foundation of Jiangsu Province of China,No.BK20151599
文摘Mounting evidence indicates that amyloid β protein（Aβ） exerts neurotoxicity by disrupting the blood-brain barrier（BBB） in Alzheimer＇s disease. Hyperoside has neuroprotective effects both in vitro and in vivo against Aβ. Our previous study found that hyperoside suppressed Aβ1-42-induced leakage of the BBB, however, the mechanism remains unclear. In this study, bEnd.3 cells were pretreated with 50, 200, or 500 μM hyperoside for 2 hours, and then exposed to Aβ1-42 for 24 hours. Cell viability was determined using 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide assay. Flow cytometry and terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling assay were used to analyze cell apoptosis. Western blot assay was carried out to analyze expression levels of Bax, Bcl-2, cytochrome c, caspase-3, caspse-8, caspase-9, caspase-12, occludin, claudin-5, zonula occludens-1, matrix metalloproteinase-2（MMP-2）, and MMP-9. Exposure to Aβ1-42 alone remarkably induced bEnd.3 cell apoptosis; increased ratios of cleaved caspase-9/caspase-9, Bax/Bcl-2, cleav ed caspase-8/caspase-8, and cleaved caspase-12/caspase-12; increased expression of cytochrome c and activity of caspase-3; diminished levels of zonula occludens-1, claudin-5, and occludin; and increased levels of MMP-2 and MMP-9. However, hyperoside pretreatment reversed these changes in a dose-dependent manner. Our findings confirm that hyperoside alleviates fibrillar Aβ1-42-induced BBB disruption, thus offering a feasible therapeutic application in Alzheimer＇s disease.
基金Supported by:Scientific and Technological Foundation of the National Administration of Traditional Chinese Medicine of China,No.02-03LP41the Scientific and Technological Key Project of Guangdong Province,No. 2006B35630007
文摘BACKGROUND： Neuronal loss, synapse mutilation, and increasing malnourished axons are pathologically related to Alzheimer＇s disease. Microtubule-associated protein 2 （MAP2） is of importance for neuronal, axonal, and dendritic generation, extension, and stabilization, as well as for the regulation of synaptic plasticity. OBJECTIVE： To investigate the antagonistic effects of natural-cerebrolysin-containing serum on beta amyloid protein 1-40 （Aβ1-40）-induced neurotoxicity from the standpoints of cell proliferation, synaptogenesis, and cytoskeleton formation （MAP2 expression）. DESIGN, TIME AND SETTING： A paralleled, controlled, neural cell, and molecular biology experiment was performed at the Institute of Integrated Chinese and Western Medicine, Shenzhen Hospital, Southern Medical University between February 2006 and April 2008. MATERIALS： PC12 cells, derived from the rat central nervous system, were purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China. A β1-40 was provided by Sigma, USA. Natural-cerebrolysin was provided by Shenzhen Institute of Integrated Chinese and Western Medicine, China. The natural-cerebrolysin was predominantly composed of Renshen （Radix Ginseng）, Tianma （Rhizoma Gastrodiae）, and Yixingye （Ginkgo Leaf） in a proportion of 1：2：2. Following conventional water extraction technology, an extract （1：20） was prepared. Each gram of extract equaled 20 grams of crude drug. In a total of 12 adult male New Zealand rabbits, six underwent intragastric administration of natural-cerebrolysin extract for 1 month to prepare natural-cerebrolysin-containing serum, and the remaining six rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： An AIzheimer＇s disease in vitro model was induced in PC12 cells using Aβ1-40. The cells were incubated with varying doses of natural-cerebrolysin-containing serum （2.5%, 5%, and 10%）. Normal blank serum-treated PC12 cells served as a blank control group. MAIN OUTCOME MEASURES： Through the use of inverted phase contrast microscope, cell morphology and neurite growth were observed, neurite length was measured, and the percentage of neurite-positive cells was calculated. Cell proliferation rate was determined by MTT assay, and MAP 2 expression was detected by fluorescent immunocytochemistry. RESULTS： Following Aβ1-40 treatments, some PC12 cells were apoptotic/dying, and only a few short neurites were observed. Following interventions with natural-cerebrolysin-containing serum, the PC12 cells proliferated, there was an increased number of neurites, and neurite length was enhanced. After middle- and high-dose natural-cerebrolysin treatments, the percentage of neurite-positive cells, as well as the average length of neurites, was significantly greater than the normal blank serum-treated PC12 cells （P 〈 0.05 or P 〈 0.01）. Compared with the blank control group, MAP2 expression in the Aβ1-40-treated PC12 cells was significantly inhibited, and the cell proliferation rate was significantly decreased （P 〈 0.01）. Following incubations with natural-cerebrolysin-containing serum, MAP2 expression and cell proliferation rate in the PC12 cells were significantly increased in a dose-dependent manner, compared with treatments with blank control serum （P 〈 0.05 or P 〈 0.01 ）. CONCLUSION： Natural-cerebrolysin exhibited antagonistic effects on neurotoxicity in Aβ1-40 induced Alzheimer＇s disease in vitro models. These effects were likely related to cell proliferation and the upregulation of intracellular MAP2 expression.
