Objective To explore the intrinsic link between neuro-endocrine-immune (NEI) network and the viscera-state.Methods Using three databases namely TCM database, Chinese pharmacy database and combination of TCM and WM database in China Academic Journals Database, the authors searched and collected NEI-related indicators published on journals for the viscera-state researches. Then a related database was established for data mining. Through analysis of association rules, analysis of the relationship among diseases, syndromes, therapeutic principles, combination of disease and syndrome, and NEI network related indicators were performed for association rules and directional network diagrams.Results Through the association analysis, the authors drew 44 directional network diagrams of high-frequency disease positions, syndromes, therapeutic principles and NEI network related indicators, and obtained 19 association rules. Kidney and liver essence research focused on HPG axis, HPA axis, and HPT axis. Spleen essence research focused on brain-gut peptide related indicators. Heart essence research focused on vascular endothelium function indicators. Pulmonary essence research focused on humeral immunity, ET and TNF-α.Conclusion It was feasible to explore the intrinsic link between NEI network and the viscera-state by using data mining. Differences among study on NEI network of five-organs systems were found, which is of great significance for researches on the essence of the viscera-state.

Objective To explore the inhibitory effects of Zibu Piyin Recipe(ZBPYR)serum on neuron apoptosis induced by tunieamyein(Tm,5 μg/ml)and its mechamsm in vitro by using sero-pharmacological method.Method Totally 12 healthy adult male SD rats(220～250 g)(SPF)were divided randomly into control group and ZBPYR group,6 in each group,then the blank and ZBPYR serum were prepared.The mouse.neuroblastoma cell line Neum2a cells were treated with Tunicamycin(Tin,an inhibitor of N-glycoslytion)to establish the endoplasmic reticulum(ER)stress model.The cells treated by ZBPYR aerum of different concentrations were interventional groups,and the cells treated by blank serum were control group.The viability of Neuro2a cells was meusurcdd by MTT assay.Flow cytometry wus applied to observe the apoptosis of Neuro2a cells.Western blotting was utilized to detect the protein expressions of two molecules,ER molecular chaperone-ucose regulated protein 78(CRP78)and transcriptional factor CCAAT/enhancer-binding protein-homologous protein(CHOP).The results were analyzed by sNK-q test.Results Compared to Tm group(cell viability 0.1673±0.0213,apoptotic rate 62.7050±1.4056),The cell viability of interventional groups(5%0.5295±0.0373,10%0.5843±0.0428,15%0.6274±0.0324)increased significantly(P<0.05);and the apoptotic rate(5%47.8733±2.8166,10%46.3366±1.2748,15%39.8833±1.0524)reduced significantly(P<0.05).The protein expressions of GRP 78(5%2.1228±0.2251,10%1.3293±0.9443,15%;15%0.0931±0.1168)and CHOP(5%1.1776±0.2927,10%0.7290±0.1708,15%0.6577±0.1883)of interventional groups reduced significantly compared with Tm group(GRP78 2.9149±0.5355;CHOP 1.6611±0.2913)P<0.05.Condusions ZBPYR serurn could increase the cell viability of Neuro2a cells treated with Tm and inhibit cell apoptosis.Thereby it may have neuroprotective effects,and the mechanism may be associated with the inhibition of ER stress and apoptosis pathway.

Objective To investigate the effects of the teaching model of team-based learning (TBL) in the course of TCM Basic Theory.MethodsTotally 61 undergraduates of Class 1-2 in Grade 2013 in Dalian Medical University are divided into 15 groups, three to five students in each group. TBL teaching model was performed in the chapter about Zang-Fu relationship in the course of TCM Basic theory. At the end of the study, the students received the questionnaire survey to know the effects of TBL teaching model.ResultsStudents discussed intensely with lively atmosphere in the class. The pass rate of individual test was 98.36%, and the excellence rate was 22.95%. The results of immediate feedback answer sheets showed that for the 6 multiple choice questions, each group answered at least 2 questions correctly for the first time, and 5 groups answered all the questions correctly for the first time with the joint efforts of group members.ConclusionTBL teaching model could promote the preview, activate atmosphere in class, improve learning efficiency, and increase the solidarity and collaboration in students.

To observe the relationship among amyloid beta-peptide (Abeta)-induced neurotoxicity, serum-inducible kinase (SNK)-spine-associated Rap guanosine triphosphatase activating protein (SPAR) pathway and N-methyl-D-aspartate receptor (NMDAR), and to explore the mechanism of the protective effect of spleen-yin nourishing recipe (Zibu Piyin Recipe, ZBPYR) in hippocampal neurons against Abeta-induced neurotoxicity.

This study was aimed to observe the effect ofZi-Bu Pi-Yin Recipe (ZBPYR) on the mRNA expressions of NMDA receptor subunits NR1, NR2A, NR2B in different brain regions of spleen-yin deficiency Alzheimer's Disease (AD) model rats. The levels of NR1, NR2A, NR2B mRNA expressions were detected by using RT-PCR method. The results showed that the levels of NR1, NR2A, NR2B mRNA expressions of AD group and spleen-yin deficiency AD group decreased significantly (P < 0.05). The levels of NR1, NR2A, NR2B mRNA expressions of ZBPYR treatment group increased significantly (P < 0.05). It was concluded that the expression levels of NMDAR mRNA in different brain regions of the ZBPYR treatment group increased significantly, which indicated that ZBPYR may up-regulate the protein expressions of NMDAR by increasing the expression levels of NMDAR mRNA, thereby to play the anti-dementia effect.

This study was aimed to explore the mechanism ofZi-Bu Pi-Yin Recipe (ZBPYR) on autophagy and endoplasmic reticulum stress (ERS) to improve spleen-yin deficiency diabetes-associated cognitive decline (DACD). Rats were randomly divided into the control (cont) group, the diabetes (DM) group, the spleen-yin deficiency (pi) group, the spleen-yin deficiency diabetes (piDM) group, and the spleen-yin deficiency diabetes + ZBPYR treatment (ZBPYR) group. The expression of microtubule-associated protein 1A/1B-light chain 3 (LC3Ⅱ), inositol-requiring enzymeα (IRE1α), c-Jun N-terminal kinase (JNK) were observed by western blot. The results showed that the expression of LC3Ⅱ in the DM group, pi group and piDM group decreased compared with the cont group (P < 0.05); and the expression of LC3Ⅱ of the ZBPYR group increased compared with the DM group and piDM group (P < 0.05). Compared with the cont group, the p-IRE1α of the DM group and piDM group, as well as p-JNK1 in the pi group and piDM group were increased (P < 0.05). The p-IRE1α and p-JNK1 of the ZBPYR group were decreased compared with the DM group and piDM group (P < 0.05). It was concluded that ZBPYR improved spleen-yin deficiency DACD by regulating autophagy and ERS.

OBJECTIVE: To determine the protective effects of nourishing spleen yin recipe (Zibu Piyin Recipe, ZBPYR), a compound traditional Chinese herbal medicine, on endoplasmic reticulum (ER) in neuronal cells responding to the stress by using sero-pharmacological method. METHODS: The mouse neuroblastoma cell line Neuro2a cells were treated with tunicamycin (Tm, an inhibitor of N-glycosylation). The ZBPYR-treated cell group was established by incubating cells with ZBPYR serum for one hour and treated with Tm. Reverse transcriptase PCR (RT-PCR) was utilized to detect the mRNA expressions from two genes after treatments, ER molecular chaperone glucose regulated protein 78 (GRP78) and transcriptional factor CCAAT/ enhancer-binding protein-homologous protein (CHOP). Lactate dehydrogenase (LDH) assay was also carried out to determine the LDH leakage from Neuro2a cells after treated with Tm and staurosporine (STS). RESULTS: The ZBPYR-treated cell group at all tested ZBPYR dosages showed significantly reduced expressions of both genes compared with Tm (5 microg/ml) treated control group (P<0.05). Therefore, ZBPYR serum inhibited the expressions of GRP78 and CHOP in mRNA level under ER stress induced by Tm. Different concentrations of ZBPYR serum pretreatment reduced the LDH leakage compared with the Tm and STS groups (P<0.05). Therefore, ZBPYR serum may inhibit the LDH leakage induced by Tm and STS. CONCLUSION: ZBPYR has neuroprotective effects. The mechanisms may be associated with inhibition of ER stress and mitochondrial dysfunction.

Objective:To apply two dimensional gel electrophoresis(2-DE) to initially analyze the protein expression in hippocampus of rats with spleen yin deficiency syndrome and to get the theory foundation for investigating the molecular mechanisms of Alzheimer's Disease and spleen yin deficiency syndrome.Methods:Rats model of spleen yin deficiency were set up by combination of improper diet,overstrain and consumption of body fluid.Qualitative changes of the proteome expression in hippocampus were compared between the model group and control group by two-dimensional gel electrophoresis.Results:The model group of spleen yin deficiency had the appearance of spleen yin deficiency,for example,irritability,increment in biting frequency,increase of rectal temperature and drinking water(P

This study was aimed to observe the changes of dendritic spine density in different regions of brain among spleen-yindeficiency dementia (SYDD) model rats, in order to investigate the effects ofZi-Bu Pi-Yin Recipe (ZBPYR) on dendritic spines. Spleen-yindeficiency (SYD) rats were modeled by classical method. And incubatedβ-Amyloid 1-40 (Aβ1-40) was injected into the hippocampus of each rat to make SYDD model, which received the administration of ZBPYR. Golgi staining was used to stain dendritic spine in different regions of brain in rat model for the observation of the amount and shape. The results showed that dendritic spine density in different regions of hippocampus and cortex in SYDD group was reduced than that of the SYD group. Compared with the dementia group and the SYDD group, the dendritic spine density in different regions of hippocampus and cortex of the SYDD + ZBPYR group was increased. Compared with the blank control group, the dendritic spine density in different regions of hippocampus and cortex in rats from the dementia group was reduced. It was concluded that there were different degrees of reducing in the dendritic spine density of different brain regions in SYDD group. ZBPYR improved the learning and memory impairment, which might be related to the maintenance of dendritic spine density in different brain regions.

This study was aimed to observe the protection ofZi-Bu Pi-Yin recipe (ZBPYR) on the cortex mitochondrial dysfunction of diabetes-associated cognitive dysfunction rats. SD rats were randomly divided into four groups, which were the control (Con) group, the diabetes (DM) group, the ZBPYR treatment (ZBPYR1) group and the ZBPYR protection (ZBPYR2) group. Morris water maze test was taken to evaluate the learning and memory ability of rats. The transmission electron microscope (TEM) was used to detect the ultrastructure and quantity changes of cortex mitochondria. Western blot was used to detect the expression of Cyto C. The results showed that compared to Con group, the learning and memory ability were decreased in the DM group (P < 0.05); the learning and memory ability of the ZBPYR1 group was improved compared to the DM group (P < 0.05); compared to the DM group, the ZBPYR2 group was significantly improved (P < 0.01). Compared with the Con group, the number of cortex mitochondria in DM group was decreased (P< 0.05), and the structure was disordered, blurred, or even completely destroyed. After ZBPYR intervention, these pathological changes were reduced obviously. And the number of mitochondria in the ZBPYR2 group was increased than that of the DM group (P < 0.05). The expression of Cyto C in cytoplasm of the DM group was significantly higher than that of the Con group (P < 0.01). After ZBPYR intervention, the expression of Cyto C was decreased. It was concluded that ZBPYR regulated the mitochondrial morphology and changes of volume in the cortex, prevented the releasing of Cyto C from mitochondria to cytoplasm, and improved the cognitive function of diabetes rats.