Molecular immunologic technique gets quick development in recent years, especially the fast development of the immuno-labelling technique, which found its indispensable status in the study of traditional Chinese medicine (TCM). With the development of the study of TCM, medical immunology connecting TCM with the Western medicine has become one of the most indispensable conjunctive subjects. Scientists can study the theories and prevention or treatment mechanisms of TCM not only at holistic and cell levels, but also at molecular and gene levels. Appropriate application of the molecular immunologic techniques according to their characteristics will play an important role in the study of Chinese integrative medicine.

AIM: To study effects of Fuyuanbunao Granules (Radix Ginseng Rubra, Fructus Crataegi, Radix Astragali, Herba Epimedii, Radix Salviae Miltiorrhizae, Rhizoma Alismatis, Rhizoma Acori Tatarinowii) on ATPase activities, MDA level in mouse brain, SOD activities in mouse blood and the quotient of brain. METHODS: On the mouse model of Alzheimer' disease toxified with AlCl 3, the effects of Fuyuanbunao Granules on Na + K + ATPase, Mg 2+ ATPase activities and MDA level in mouse brain, SOD activities in mouse blood were evaluated with spectroscopic analysis, and the weight of brain was measured with analytical balance. RESULTS: Compared with Alzheimer'disease model team, Fuyuanbunao Granules enhance the activities of Na +、K + ATPase, Mg 2+ ATPase, decease MDA level in the mouse brain and enhance the activities of SOD in blood. It also increases the weight of brain. CONCLUSION: It indicates that enhancing the activity of ATPase and deceasing peroxide level may be one of the mechanism for Fuyuanbunao Granules to prevent and cure Alzheimer' disease.

Objective To compare the outcomes of patients starting peritoneal dialysis (PD)within two weeks and more than two weeks after catheter implantation.Methods All the patients undergoing Tenckhoff catheter implantation and initiating PD in Renji Hospital from January 2001 to December 2010 were enrolled in the study.Patients started PD within 2 weeks after catheter insertion were defined as urgent group,and those started PD 2 weeks later were defined as planned group.Kaplan-Meier curves and Log-rank tests were used to compare outcomes between two groups.Results Among 657 patients in this study,median break-in period was 6 days of 469 (71.4％)patients in urgent group and 26 days of 188 (28.6％) patients in planned group.Compared to planned group,patients of urgent group were younger [(52.6± 17.3) vs (56.1± 15.3) year,P =0.017],had less eGFR [(5.36±2.03) vs (6.50±2.50) ml· min-1 · (1.73 m2)-1,P ＜ 0.01],lower serum albumin [(34.0±5.7) vs (36.2±5.9) g/L,P ＜ 0.01] and hemoglobin [(76.9± 18.8) vs (80.8 ± 17.9) g/L,P =0.018],and higher phosphate [(2.19±0.67) vs (1.98±0.52) mmol/L,P＜ 0.01].Urgent group presented more catheter dysfunctions needed to transfer to hemodialysis (2.1％ vs 0％,P =0.044).The 1-,2-,3-and 5-year technique survival rates of urgent and planned group were 94％ vs 98％,92％ vs 94％,90％ vs 92％and 86％ vs 85％ respectively.There was no significant difference in technique survival (Log-rank =1.536,P =0.22) and peritonitis-free survival (Log-rank =0.035,P =0.85) between two groups.The 1-,2-,3-and 5-year patient survival rates of urgent and planned group were 90％ vs 95％,81％ vs 90％,74％ vs 79％ and 67％ vs 74％ respectively,and no significant difference was found (Log-rank =2.364,P =0.12).Conclusions Although patients needing urgent initial PD have poorer residual renal function and nutritional condition compared to those of planned initial PD,their outcomes are similar.Peritoneal dialysis may be a feasible and safe dialysis modality for patients who need urgent start.

Objective To investigate the impact of calcium phosphate crystals induced by uremic serum on calcification of human aortic smooth muscle cells (HASMCs).Methods Uremic serum was incubated at 37℃ for 3 days.Calcium phosphate crystals and uremic supernatant were isolated from uremic serum by uhracentrifugation.Scanning electron microscope (SEM) and energy dispersive X-ray spectroscopy (EDX) were performed for analysis of morphological and chemical characteristics of the crystals.HASMCs were treated in vitro with control medium,uremic serummedium,calcium phosphate crystals-medium and uremic supernatant-medium.Calcification was visualizcd by Alizarin red staining.Calcium loads in cells were quantified by o-cresolphthalein complexone method.Gene expression of bone morphogenetic protein-2 (BMP-2),osteopontin (OPN) and core-binding factor α1 (Cbfα1),alkaline phosphate (ALP) and matrix gamma carboxyglutamic acid protein (MGP) were quantified by real-time PCR.Cbfα1,OPN and BMP-2 protein levels were determined by Western blotting or ELISA.Results Calcium phosphate crystals which induced by uremic serum displayed laminated shapes containing crystallized needle-like projections and ranged from 30-500 nm,with Ca/P ratios of 1.41 ±0.05.Compared with the cells in control group,uremic serum induced HASMCs calcification,increased calcium loads (P ＜ 0.05),up-regulated BMP-2,OPN,Cbfα1 mRNA and protein expression (all P＜ 0.01).Similar to uremic serum,calcium phosphate crystals also induced HASMCs calcification,increased calcium loads (P＜0.05),and up-regulated BMP-2,OPN,Cbfα1 mRNA and protein expression (all P ＜ 0.01).However,there was no significant difference between HASMCs growing in uremic supernatant and control medium in calcium loads or the expression levels of these osteogenic proteins (P ＞ 0.05).Conclusions Calcium phosphate crystals induced by uremic serum promote HASMCs calcification,which might be one of the mechanisms of uremic vascular calcification.

Objective To investigate the mechanism of Guiqiyiyuan Ointment (GO) for preventing and treating 12C6+ beam radiation induced lung and kidney bystander effect to provide a new strategy for prevention and treatment of clinical radiation injury. Methods Sixty healthy male Wistar rats were randomly and equally divided into 3 groups: NC group, SR group (simple radiation 2ml/kg), GO group(GO 2ml/kg intragastric administration for 7 days). The right side of the lung was modeled by 12C6+beam radiation. After modeling, the rats were killed at 48h. The left lung, left and right kidney tissues were taken from the rats. The DNA methylation rate was detected by ELISA assay, pathological changes were observed by HE staining, and the expressions of Dnmt1, Dnmt3a and Dnmt3b were detected by immunohistochemistry. Results Compared with the NC group, the level of DNA methylation was decreased significantly (P<0.01), the left lung showed inflammation, no abnormal finding was seen in the left and right kidneys, and the expressions of Dnmt1, Dnmt3a and Dnmt3b were significantly increased in the SR group (P<0.01). Compared with the SR group, the level of DNA methylation was increased significantly (P<0.01), the left lung inflammation became better, and the expressions of Dnmt1, Dnmt3a and Dnmt3b were significantly decreased in the GO group (P<0.01). Dnmt1, Dnmt3a and Dnmt3b proteins were expressed in the cytoplasms of bronchial and renal tubular epithelial cells in all the groups. The NC group presented as light brown-brown staining, showing a weak positive expression, the SR group as brown-brown staining, showing astrong positive expression, and the GO group as light brown-brown staining, showing a moderate positive expression. Conclusion The GO can reduce the bystander effect caused by 12C6+ beam radiation, and its mechanism is related to improving the level of DNA methylation.

Objective To investigate the role of BMP-2/Smad signaling pathway in the osteogenic differentiation of human aortic smooth muscle cells (HASMCs) caused by hyperphosphatemia -induced calcium phosphate (CaP) crystals.Methods High-phosphate medium was incubated at 37℃ for 3 days.CaP crystals and supernatant were isolated by ultracentrifugation.Scanning electron microscope and energy dispersive X-ray spectroscopy were performed for analysis of physicochemical characteristics of CaP crystals.HASMCs were cultured in vitro,and divided into high-phosphate,control,crystals and supernatant groups.Calcification was visualized by Alizarin red staining.Calcium loads in cells were quantified by o-cresolphthalein complexone method.Protein expression of bone morphogenetic protein-2 (BMP-2),Runt-related transcription factor 2 (RUNX2),osteopontin (OPN),phospho-Smad1/5/9 (p-Smad1/5/9) were quantified by Western blotting.After knockdowns of BMP-2 and Smad1 with small hairpin RNA (shRNA) interfering respectively in HASMCs,protein expressions were measured by Western blotting.Results High-phosphate medium induced the formation of CaP crystals.Compared with the cells in control group,CaP crystals significantly induced HASMCs calcification,increased calcium loads and up-regulated the levels of BMP-2,RUNX2 and OPN proteins (all P ＜ 0.05).After the addition of CaP crystals into HASMCs,the level of p-Smad 1/5/9 protein peaked at 30 min (P ＜ 0.05).After BMP-2 was knocked down in HASMCs,the expression of p-Smad1 caused by CaP crystals was blocked completely,and the expressions of RUNX2 and OPN caused by CaP crystals were reduced significantly (all P ＜ 0.05).After Smad1 was knocked down in HASMCs,the expressions of RUNX2 and OPN caused by CaP crystals were decreased significantly (all P ＜ 0.05).Conclusions Hyperphosphatemia-induced CaP crystals promoted osteogenic differentiation of HASMCs through the BMP-2/Smad signaling pathway.

Objective To predict the target and molecular mechanism of Huayu Xiaopi decoction in the intervention of Precancerous lesions of gastric cancer(PLGC)based on network pharmacology and molecular docking technology,and to conduct experimental verification.Methods A total of 60 SPF SD male rats were randomly selected as blank control,and the other rats were replicated in PLGC model.After successful modeling,the rats were randomly divided into model group,folic acid group(2 mg·kg-1·d-1),Huayu Xiaopi decoction high,medium and low dose groups(24.8,12.4,6.2 g·kg-1·d-1),which were continuously administered for 90 days.The body mass and food intake of rats at 3 h were recorded,and the gastric histopathology was observed by HE staining.Network pharmacology and molecular docking techniques were used to predict the potential targets of Huayu Xiaopi decoction in PLGC intervention,and the core targets were verified by Western blot technique.Results Compared with the blank group,the body mass and 3 h food intake of rats in the model group were significantly decreased(P<0.05),the gastric mucosa of rats was significantly thinner,the glands were significantly reduced and disordered,and the intestinal metaplasia goblet cells and a large number of inflammatory cells were visible in some areas.Compared with the model group,the body mass and 3 h food intake of rats in each administration group were improved to varying degrees.Huayu Xiaopi Decoction improved significantly in medium and high doses(P<0.05),the gastric mucosa was repaired in different degrees,the glandular arrangement tended to be orderly,and the inflammatory cells in the interstitial were gradually reduced.The results of network pharmacology and molecular docking showed that TP53,JUN and MAPK3/1(ERK1/2)were the core targets of Huayu Xiaopi decoction in the intervention of PLGC.Molecular biological detection results showed that compared with blank group,the protein phosphorylation levels of TP53,c-Jun and ERK1/2 in gastric tissue of model group were significantly increased(P<0.05).Compared with model group,the protein phosphorylation levels of TP53,c-Jun and ERK1/2 in gastric tissue of rats in all administration groups were decreased to different degrees,and significantly decreased in Huayu Xiaopi decoction high-dose and medium-dose groups(P<0.05).Conclusion Huayu Xiaopi Decoction can significantly improve the survival condition of PLGC rats and promote gastric mucosal repair,the specific mechanism of which may be related to the decrease of ERK1/2,c-Jun and TP53 protein phosphorylation levels in gastric tissue of PLGC rats,and then regulate the downstream signaling molecular response.

ObjectiveTo explore the intervention mechanism of Xiangsha Liujunzi Tang in rats with functional dyspepsia （FD） based on the Ras homolog gene family member A （RhoA）/Rho-associated coiled-coil containing protein kinase 2 （ROCK2）/Myosin phosphatase target Subunit 1 （MYPT1） pathway. MethodSixty male SD suckling rats in SPF grades were randomly divided into blank group （n=10） and model group （n=50）. The comprehensive modeling method （gavage administration of iodoacetamide+exhaustion of swimming+disturbance of hunger and satiety） was used to replicate the rat model of FD. After successful replication of the model， the rats in the model group were randomly divided into model group， mosapride group， and high， middle， and low-dose Xiangsha Liujunzi Tang groups， with 10 rats in each group. Rats in the blank group and model group were given 10 mL kg^-1·d^-1 normal saline， those in the mosapride group were given 1.35 mg·kg^-1·d^-1 mosapride， and those in the high， middle， and low-dose Xiangsha Liujunzi Tang groups were given 12， 6， and 3 g·kg^-1·d^-1 Xiangsha Liujunzi Tang， respectively. The intervention lasted 14 days. The general living conditions of rats were observed before and after modeling and administration， and the 3-hour food intake and body mass of rats were measured. After intervention， the intestinal propulsion rate of rats was measured， and the pathological changes in the gastric tissue were observed by hematoxylin-eosin （HE） staining. The content of choline acetyl transferase （ChAT） and vasoactive intestinal peptide （VIP） in the medulla oblongata and gastric tissue homogenate was determined by enzyme-linked immunosorbent assay （ELISA）， the distribution of adenosine triphosphate （ATP） enzyme in gastric antrum smooth muscle was observed by frozen section staining， and the protein expression levels of RhoA， ROCK2， and phosphorylated-myosin phosphatase target subunit 1 （p-MYPT1） in the gastric tissue were detected by Western blot. ResultCompared with the blank group， the model group had withered hair， lazy movement， slow action， poor general living condition， lower 3-hour food intake， body mass， and lower intestinal propulsion rate （P<0.05）， whereas no obvious abnormality in gastric histopathology. In the model group， the content of ChAT in the medulla oblongata and gastric tissue decreased， the content of VIP in gastric tissue increased， the distribution of ATP enzyme in gastric antrum smooth muscle decreased significantly， and the protein expression levels of RhoA， ROCK2， and p-MYPT1 in the gastric tissue decreased significantly （P<0.05）. As compared with the model group， the general living condition of rats in each intervention group was significantly improved， and the 3-hour food intake， body mass， and intestinal propulsion rate were significantly increased （P<0.05）. There was no significant difference in gastric pathology in the intervention groups. The content of ChAT in the medulla oblongata and gastric tissue increased significantly， the content of VIP in the gastric tissue decreased， the distribution of ATP enzyme in gastric antrum smooth muscle increased significantly， and the protein expression levels of RhoA， ROCK2， and p-MYPT1 in the gastric tissue increased significantly （P<0.05）. The intervention effect of Xiangsha Liujunzi Tang group on the above indexes was dose-dependent. ConclusionXiangsha Liujunzi Tang can effectively improve the general living condition and gastric motility of rats with FD， and its specific mechanism may be related to the activation of the RhoA/ROCK2/MYPT1 pathway in the gastric tissue to regulate smooth muscle relaxation and contraction and promote gastric motility.