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 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.

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.

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.

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 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

Objective To clarify the pathogenesis of spleen yin deficiency diabetes-associated cognitive decline (DACD) and mechanism of Zinu Piyin Recipe (ZBPYR) by observing the expression of phosphorylated RNA-dependent protein kinase-like endoplasmic reticulum kinase (PERK), eukaryotic initiation factor 2α subunit (eIF2α), p-eIF2α, glucose-regulated protein 78 (GRP78) in cerebral cortex of spleen yin deficiency diabetes mellitus rats. Methods The rats were randomly divided into control group, diabetes mellitus group, spleen yin deficiency group, spleen yin deficiency diabetes mellitus group (disease-syndrome group) and spleen yin deficiency diabetes mellitus+ZBPYR group (treatment group). Type 2 diabetes mellitus models were established by high-fat food feeding for 4 weeks and low dose STZ intraperitoneal injection. Then the classical compound method was used to construct spleen yin deficiency rats model by improper diet, over exertion and yin fluids exhaustion. The reatment group was given ZBPYR and other groups were given equal volume of normal saline for 15 days, then cerebral cortex was obtained. The expression of p-PERK, p-eIF2α, eIF2α and GRP78 were observed by Western Blot and RT-PCR. Results The protein expression of p-PERK, p-eIF2α, and mRNA expression of GRP78 of diabetes mellitus group, spleen yin deficiency group and disease-syndrome group was enhanced compared with control group (P<0.05). GRP78 protein expression of diabetes mellitus group, disease-syndrome group was increased compared with control group (P<0.05). The protein expression of p-PERK, p-eIF2α, GRP78 and mRNA expression of GRP78 of treatment group was decreased compared with diabetes mellitus group and disease-syndrome group (P<0.05). Conclusion Endoplasmic reticulum stress is involved in spleen yin deficiency DACD. ZBPYR improves learning and memory ability by reducing endoplasmic reticulum stress.

Objective To explore the mechanism of Zibu Piyin Recipe (ZBPYR) on spleen yin deficiency diabetes-associated cognitive disorder (DACD). Methods The rats were randomly divided into control group, diabetes mellitus (DM) group, spleen yin deficiency group, spleen yin deficiency DM group and spleen yin deficiency DM+ZBPYR group (treatment group). Type 2 DM models were established by high-fat food feeding and low dose STZ intraperitoneal injection for 4 weeks. Then the classical compound method was used to construct spleen yin deficiency rat models by improper diet, over exertion and yin fluids exhaustion. The treatment group was given ZBPYR by gavage for 15 days, and the other groups were given the same amount of normal saline. Then cerebral cortex, hippocampus, stomach and liver were obtained and the changes of protein expression of PDHE1α in them were observed by Western Blot. Results The protein expression of PDHE1αin cortex of DM group and spleen yin deficiency DM group were lower than control group (P<0.05). PDHE1α expression of treatment group in cortex and stomach increased more significantly than spleen yin deficiency DM group (P<0.05). The expression of PDHE1α protein showed no significant difference among all groups in hippocampus and liver. Conclusion ZBPYR improved spleen yin deficiency DACD by regulating PDHE1αin cortex and stomach.