OBJECTIVE: To investigate the effects of huperzine A in ameliorating acute hypobaric hypoxic-induced spatial memory deficits and in relieving oxidative stress injury in rat brain. METHODS: A total of 48 rats were randomly divided into four groups, the champaign group (control group), champaign + huperzine A group (0.1 mg · kg-1), high altitude group (simulated 6000 m plateau) and high altitude + huperzine A group. The Morris water maze learning and memory test, the concentration of GSH, MDA as well as the activities of CAT, SOD and LDH in hippocampus were measured and compared. RESULTS: Compared with those of high altitude group, escape latency of high altitude + huperzine A group was significantly shorter (P < 0.05), platform crossings within 60 s was significantly more (P < 0.05), time spent in target was significantly longer (P < 0.05), GSH content, SOD and CAT activity in hippocampal tissue were significantly higher (P < 0.05), MDA and LDH activity in hippocampal tissue was significantly lower (P < 0.05), and all these changes had no significantly difference as compared with the champaign group. CONCLUSION: Huperzine A treatment has protective effects against acute hypobaric hypoxic-induced oxidative stress injury in rat brain, and ameliorates spatial memory deficits in rats. Copyright 2012 by the Chinese Pharmaceutical Association.

Purpose:To study the expression of CD44v6 protein in breast carcinoma and its prognostic significance. Methods:100 cases of formalin-fixed paraffin-embeded female breast invasive ductal carcinoma tissues were retrospectively analyzed using EnVision~(TM) immunohistochemical method with the monoclonal antibody CD44v6.Statistical analysis was based on the Log-rank test and Cox analysis.Results:Sixty-six of 100 cancer tissues expressed CD44v6.Positive staining was mainly on the cell membranes.There was significant correlation between CD44v6 immunoreactivity and lymph node me- tastasis and TNM stage.The 5-and 10-year survival rates were 82.76% and 78.37% of patients with CD44v6 low-expres- sion tumors,and 64.1% and 49.88% of those with CD44v6 high-expression tumors,respectively;the difference between the two groups of patients was significant(P

Objective To observe the expression of matrix metalloproteinases(MMP)-1,MMP-3 and iNOS in cartilage of experimental rabbit osteoarthritis at different time intervals after anterior cruciate ligament transection(ACLT)operation.The aim of this study is to provide the theoritical evidence for using ACLT rabbit model in Osteoarthritis(OA)research.Methods Unilateral ACLT was performed on 27 randomly selected while rabbits and underwent unilateral arthrotomy was performed on the other 9 white rabbits as the control group.Nine randomly selected white rabbits in experimental group were killed and 3 white rabbits in the control group at 4th,8th and 12th week respectively.Cartilage degradation of femoral condyles was evaluated macr-oscopically,mRNA expression level and protein expression level of MMP-1,MMP-3 and iNOS was measured by reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemistry respectively.Results Forepart OA cartilage degradation was observed at the 4th week and became more severe at the 8th week after ACLF.Afterpart cartilage degradation was evident at the 12th week after ACLT while cartilage still remained normal in the control group,mRNA expression level and protein expression level of MMP-1.MMP-3 and iNOS were increased at the 4th week and became higher gradually at the 8th,12th week after ACLT compared with the control group.Expression distribution of MMP-1,MMP-3 and iNOS bad different patterns respectively.Conclusion It is suggested that the process of OA cartilage degradation can be simulated by ACLT model and MMP-1,MMP-3 and iNOS may be good markers in therapeutical research of OA.

OBJECTIVETo establish discriminant functions of diarrhea-predominant irritable bowel syndrome (IBS-D) by studying it from quantitative diagnosis angle, hoping to reduce interference of subjective factors in diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

METHODSA Chinese medical clinical epidemiological survey was carried out in 439 IBS-D patients using Clinical Information Collection Table of IBS. Initial syndromes were obtained by cluster analysis. They were analyzed using step-by-step discrimination by taking information of four Chinese medical diagnostic methods and serum brain-gut peptides (BGP) as variables.

RESULTSClustering results were Gan stagnation Pi deficiency syndrome (GSPDS), Pi-Wei weakness syndrome (PWWS), Gan stagnation qi stasis syndrome (GSQSS), Pi-Shen yang deficiency syndrome (PSYDS), Pi-Wei damp-heat syndrome (PWDHS), cold-damp disturbing Pi syndrome (CDDPS). Of them, GSPDS was mostly often seen with effective percentage of 34. 2%, while CDDPS was the least often seen with effective percentage of 5.5%. A total of 5 discriminant functions for GSPDS, PWWS, GSQSS, PSYDS, and PWDHS were obtained by step-by-step dis- crimination method. The retrospective misjudgment rate was 4.1% (16/390), while the cross-validation misjudgment rate was 15.4% (60/390).

CONCLUSIONThe establishment of discriminant functions is of value in objectively diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

Alarmins ; Brain ; Cluster Analysis ; Diarrhea ; classification ; diagnosis ; Hot Temperature ; Humans ; Irritable Bowel Syndrome ; classification ; diagnosis ; Medicine, Chinese Traditional ; Qi ; Retrospective Studies ; Surveys and Questionnaires ; Yang Deficiency

To observe the effects of Danshen-containing serum on SuFu and DYRK2 expression in the HSCs stimulated by leptin. SD rats (n = 60) were used to make danshen-containing serum by gastric perfusion for ten days with Danshen water decoction, normal saline and colchicine. The HSCs that were cultured in vitro would be stimulated for 24 hours by leptin (100 μg x L(-1)) except blank control group, after being intervened, the drug serum in each group would be cultured at 37 degrees C in 5% incubator. The cells would be collected after 24 hours, then the effects of danshen-containing serum on the proliferation of HSCs were detected by MTT, the expression of SuFu mRNA and DYRK2 mRNA were detected by RT-PCR, the expression of SuFu and DYRK2 proteins were tested by Western blot. Compared with blank control group, the expression of DYRK2 mRNA and DYRK2 proteins were enhanced obviously after stimulated the HSCs of rats by leptin (P < 0.01), but the expression of SuFu mRNA and SuFu proteins were decreased significantly (P < 0.01). Compared with the model group, after cyclopamine group (Hh pathway inhibitor), Danshen-containing serum and colchicine were interfered, the expression of DYRK2 mRNA and DYRK2 proteins were decreased clearly (P < 0.01), but the expression of SuFu mRNA and SuFu proteins were increased significantly (P < 0.01 or P < 0.05). Compared with model group, adding purmorphamine (Hh pathway agonist) to model group and making it activate could increase the expression of DYRK2 mRNA and DYRK2 proteins, but the expression of SuFu mRNA and SuFu proteins were decreased significantly (P < 0.01). Compared with the model group, using the Danshen-containing serum to interfere the purmorphamine group could make the expression of DYRK2 mRNA and DYRK2 proteins decrease and the expression of SuFu mRNA and SuFu proteins increase significantly (P < 0.01). Danshen-containing serum would inhibition the activation and increment of HSCs by interfering the expression of SuFu and DYRK2 which were induced by leptin.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatic Stellate Cells ; drug effects ; metabolism ; Humans ; Liver Cirrhosis ; drug therapy ; genetics ; metabolism ; Male ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Repressor Proteins ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo explore the scavenging action of tenuigenin (TEN) on intracerebral amyloid β protein (Aβ) aggregation and the abnormal phosphorylated tau protein and its mechanism in Alzheimer's disease (AD) rats' brain.

METHODSAβ1-40 was injected into the right CA1 region hippocampus to establish the AD model. Successfully modeled rats were divided into the model group, the low, middle, high TEN group. Rats were administered with TEN (18.5, 37.0, 74.0 mg/kg) by gastrogavage. Besides, a sham-operation group was set up. Expression levels of Aβ1-40 and Tau p-Ser262 were detected by immunohistochemistry. Expression levels of ubiquitin (Ub) and Ub-protein ligase E3 were measured by Western blotting.The content of 26S proteasome was detected by ELISA.

RESULTSImmunohistochemical results showed that the number of Aβ and Tau p-Ser262 positively reacted neurons significantly increased in model group, when compared with the sham-operation group (P < 0.01). Results of Western blot showed expression levels of ubiquitinated protein were up-regulated and those of Ub-protein ligase E3 were down-regulated in the model group (P < 0.01). ELISA results showed that the content of 26S proteasome significantly decreased in AD rats' brain (P < 0.01). Compared with the model group, expression levels of Aβ1-40, Tau p-Ser262, and Ub significantly decreased; expression levels of Ub-protein ligase E3 apparently increased; the content of 26S proteasome significantly increased in each TEN treatment group (P < 0.05, P < 0.01). Best effect was shown in 37.0 mg/kg and 74.0 mg/kg TEN groups.

CONCLUSIONSUb proteasome pathway (UPP) participated in the occurrence of AD. TEN could obviously reduce intracere- bral Aβ1-40 accumulation and abnormal tau phosphorylation.

Alzheimer Disease ; metabolism ; Amyloid beta-Peptides ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hippocampus ; metabolism ; Neurons ; metabolism ; Phosphorylation ; Proteasome Endopeptidase Complex ; metabolism ; Rats ; Ubiquitin-Protein Ligases ; metabolism ; Ubiquitins

Objective: To observe the clinical efficacy difference in treating irritable bowel syndrome-diarrhea (IBS-D) of liver-qi stagnation and spleen-deficiency pattern with different treatment protocols, and the effects on serum levels of 5-hydroxytryptamine (5-HT), substance P (SP) and vasoactive intestinal peptide (VIP), for unveiling the mechanism of intradermal needle therapy plus pinaverium bromide in treating IBS-D. Methods: A total of 123 IBS-D patients were divided into an observation group, a Western medication group and an integrated Western and Chinese medication group using the random number table method, with 41 cases in each group. The Western medication group was given oral pinaverium bromide, 50 mg each time and 3 times a day. The integrated Western and Chinese medication group was given additional Chinese herbal medicine Tong Xie Yao Fang, one dose each day. The observation group was given additional intradermal needle therapy on the basis of the Western medication group. The whole intervention lasted for 6 weeks. Before and after treatment, the scores of gastrointestinal symptoms, traditional Chinese medicine (TCM) symptoms, irritable bowel syndrome (IBS) symptom severity scale (IBS-SSS) and IBS quality of life (IBS-QOL) questionnaire, as well as the serum levels of 5-HT, SP and VIP were observed. The clinical efficacy was estimated. Results: The total effective rate was 92.7％ in the observation group, 68.3％ in the Western medication group and 78.1％ in the integrated Western and Chinese medication group. The total effective rate was higher in the observation group than in the other two groups, and higher in the integrated Western and Chinese medication group than in the Western medication group, showing statistical significance (all P<0.05). After treatment, the scores of gastrointestinal symptoms, TCM symptoms and IBS-SSS showed significant decreases in the three groups, presenting statistical significance compared with the baseline (all P<0.05); the scores of gastrointestinal symptoms, TCM symptoms and IBS-SSS were notably lower in the observation group than in the other two groups (all P<0.05), and lower in the integrated Western and Chinese medication group than in the Western medication group (all P<0.05). After treatment, the eight component scores of IBS-QOL showed significant increases in the three groups compared with the baseline (all P<0.05); the eight component scores in IBS-QOL were significantly higher in the observation group than in the other two groups (all P<0.05), and higher in the integrated Western and Chinese medication group than in the Western medication group (all P<0.05). After treatment, the serum levels of 5-HT, SP and VIP decreased markedly in the three groups compared with the baseline (all P<0.05); the serum levels of 5-HT, SP and VIP were significantly lower in the observation group than in the other two groups (all P<0.05), and lower in the integrated Western and Chinese medication group than in the Western medication group (P<0.05). Conclusion: Treatment with intradermal needle therapy plus pinaverium bromide results in significant improvements in the gastrointestinal symptoms and quality of life in patients with IBS-D of liver-qi stagnation and spleen deficiency pattern, and effectively regulates the gastrointestinal hormone production.

Objective To investigate the effects of huperzine A on ameliorating acute hypobaric hypoxic-induced spatial learning and memory deficits, and on relieving the apoptosis of hippocampal neurons in rats. Methods Forty-eight SD rats were randomly divided into four groups (12 each): the champaign (plain) group (control group), champaign+huperzine A group, high altitude group (simulated 6000m plateau) and high altitude+huperzine A group. One day before the decompression simulation experiment, rats in huperzine A-treated groups were given intragastrically with huperzine A suspension (10mg/ml) in a dose of 0.1mg/kg. The spatial learning and memory performance of rats in each group were tested by Morris Water Maze. The apoptosis of hippocampal neurons was determined by TUNEL. The expression of pro-apoptotic proteins (Bax) and anti-apoptotic protein (Bcl- 2) of hippocampus tissues were evaluated by Western blotting. Results Compared with those in high altitude group, significantly shortened escape latency (P<0.05), more platform crossing within 60s (P<0.05), longer retention time in target (P<0.05), lower rate of hippocampal neurons apoptosis (P<0.05), down-regulated expression of Bax (P<0.05) and up-regulated expression of Bcl- 2 (P<0.05) in the hippocampus tissues were found in the high altitude+huperzine A group. However, no significant difference in the above mentioned findings was found between high altitude+huperzine A group and champaign control group. Conclusion Huperzine A treatment may have a protective effect against acute hypobaric hypoxic-induced apoptosis of hippocampal neurons in rats, and it ameliorates spatial learning and memory deficits in rats.

Teaching method for basal experiment, comprehensive experiment, design experiment and teach- ing practice in food microiological analysis were elaborated completely, and design experimental teaching was discussed stress. At the same time, Through introducing various experience of the design experiment teaching, resolvent and way of thinking against problem meeted in design experiment teaching were put forward.