Network pharmacology is an emerging discipline based on the Disease-Gene-Drug multilevel network. And it has been used to forecast the drug targets and improve the efficiency of drug discovery. Its research ideas are similar to the overall efficacy of traditional Chinese medicine (TCM), which attracts more and more medical re-searchers to look for the joint point of TCM and network pharmacology. A series of approaches on disease-related genes, predicting the information of target and active ingredients of TCM emerge. In this paper, the network pharma-cology research tools, databases and their applications were summarized and introduced. This paper also proposed scientific strategies to separate active ingredients of TCM using network pharmacology, so as to improve the efficiency and speed of finding active ingredients of TCM.

Objective To evaluate the quality of medicinal materials of Lignum Dalbergiae Odoriferae in Chinese herbal medicine market. Methods Eighteen batches of commercial medicinal materials of Lignum Dalbergiae Odoriferae were identified and analyzed by macroscopical identification, thin layer chromatography (TLC), and volatile oil assay according to the Chinese Pharmacopoeia published in 2010. Visible spectrometry was used to determine the content of total flavonoids from the qualified samples. And the gas chromatography was applied to evaluate the content of nerolidol from volatile oils of the qualified samples. Results Only 33.3%of the samples met the standard of Chinese Pharmacopoeia. The total flavonoid content of the qualified samples was in the range of 21.6-29.0 mg/g. The content of nerolidol from volatile oils of the qualified samples was in the range of 294-574 mg/g. Conclusion At present, the quality of medicinal materials of Lignum Dalbergiae Odoriferae in Chinese herbal medicine market and in clinic varies greatly, and adulterants and inferior are common. The contents of chemical components in different batches of samples are significant different.

This study aimed to construct recombinant adenovirus expressing Epstein-Barr nuclear antigen 1 (EBNA1) against nasopharyngeal carcinoma (NPC). The C-terminal region fragment of the ebna1 gene of Epstein-Barr virus was amplified from the standard strain B95-8 by polymerase chain reaction (PCR). The gene fragment was inserted into the pDC316 shuttle plasmid using the EcoRI and BgIII restriction enzyme sites. The pDC316-ebna1 shuttle plasmid and pBHG helper plasmid were cotransfected into HEK293 cells after sequencing. The soluble protein was extracted from HEK293 cells, which caused apparent cytopathic effects. The transcription and expression of the ebna1 gene were confirmed using flow cytometry and Western blotting. rAd-ebna1 titers were measured by the TCID50. rAd-ebna1 was injected into BALB/c mice at a dose of 2 x 10(8) VP per mouse, EBNA1 epitope-specific responses were measured at 1st, 2nd, 4th and 8th weeks post-immunization. The target fragment of ebna1 (939 bp) was obtained by PCR, and was in consensus with the sequence from the standard strain B95-8. Cytopathic effects were observed after the pDC316-ebna1 shuttle plasmid and pBHG helper plasmid were cotransfected into HEK293 cells. rAd-ebna1 was successfully recombined in HEK293 cells. EBNA1 protein was detected in HEK293 cells, rAd-ebna1 titers reached 10(8) TCID50/mL. Specific responses to CD4+ epitopes of EBNA1 were detected in the immunized mice. In conclusion, rAd-ebna1 was successfully constructed and induced specific responses to CD4+ epitopes of EBNA1 in immunized mice.
Adenoviridae ; genetics ; immunology ; Animals ; CD4-Positive T-Lymphocytes ; immunology ; virology ; Epstein-Barr Virus Infections ; immunology ; prevention & control ; virology ; Epstein-Barr Virus Nuclear Antigens ; administration & dosage ; genetics ; immunology ; Genetic Vectors ; genetics ; immunology ; Herpesvirus 4, Human ; genetics ; immunology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Viral Proteins ; administration & dosage ; genetics ; immunology

Objective To assess the risk factors of the in-hospital mortality of acute type A aortic dissection after operation. Method From January 2003 to June 2008,185 patients, 144 males and 41 females, with acute type A aortic dissection operated on were enrolled. The average age of patients was (49.46 ± 11.04 ) years old.The patients' demographics, history, clinical features, and some laboratory examinations were reviewed. Univariate and multivariate analysis followed by logistic regression analysis were carried out to identify the predictors of inhospital mortality. Results The in-hospital mortality rate was 9.1%. The results of univariate and multivariate analyses as follows: pre-operation positive neurological symptom (Univariate OR = 5.084,95%CI:1.792 -14.426, P = 0.002; Multivariate OR = 5.538,95%CI: 1.834 - 16.721, P = 0.002, respectively), hypotension (Univariate OR = 6.986,95%CI:1.510- 32.323,P =0.013; multivariate OR = 1.998,95%CI:0.315-12.679,P = 0.463, respectively) and renal failure (Univariate OR = 3.594,95%CI:1.237 - 10.438,P =0.019; Multivariate OR = 3.254,95%CI:1.034- 10.242, P= 0.044, respectively). Conclusions There are two predictors, pre-operation positive neurological symptom and renal failure, of pre-hospital mortality found in current analyses. Our results may improve the regimen made by cardiac surgeons and emergency doctors so as to help patients and their relatives to make correct decision.

OBJECTIVETo observe the effect of Migu capsule and Strengthening Spleen prescriptions on the expression of vitamin D receptor on small intestine and calf muscle of the rat, while observing whether the Chinese medicine complex prescriptions of different effect such as invigorating the kidney and strengthening the spleen had selectivity to expression of the VDR mRNA.

METHODSCopy the model of osteoporosis by ovariectomy operation, the rats were randomly divided into four groups, pseudo-resection group, model group, Migu capsule group and strengthening Spleen prescriptions group. Drug delivery 12 weeks later from operation, stomach lavage last for 12 weeks. Then, to observe the effect of medicine on the rats' bone mineral density, uterus weight, calf muscle/weight, and the VDR mRNA in small intestine and calf muscle through RT-PCR.

RESULTSMigu capsule enhanced bone mineral density (P<0.05), raised calf muscle/weight (P<0.05), up-regulated expression of calf muscle VDR mRNA in the small intestine (P<0.01). Strengthening Spleen prescriptions remained bone mineral density (P>0.05), up-regulated expression of calf muscle VDR mRNA (P<0.01), there was no obvious difference in uterus weight in Migu capsule group and Strengthening Spleen prescription group.

CONCLUSIONMigu capsule and Strengthening Spleen prescriptions can cure osteoporosis by improving the expression of calf muscle VDR mRNA in the small intestine.

Animals ; Bone Density ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Regulation ; drug effects ; Intestine, Small ; drug effects ; metabolism ; Muscle, Skeletal ; drug effects ; metabolism ; Osteoporosis ; metabolism ; prevention & control ; Ovariectomy ; Prescriptions ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Calcitriol ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Spleen ; drug effects ; metabolism

Endoplasmic reticulum (ER) stress occurs in macrophage-rich areas of advanced atherosclerotic lesions and contributes to macrophage apoptosis and subsequent plaque necrosis. The purpose of the present study was to investigate the effects of caveolin-1 (Cav-1) on ER stress-induced apoptosis in cultured macrophages and the underlying mechanisms. RAW264.7 cells were incubated with thapsigargin (TG) to establish ER stress model. And Cav-1 expression was detected by Western blot. After being pretreated with filipin(III), a caveolae inhibitor, RAW264.7 cells were assayed with flow cytometry and confocal laser scanning microscopy to detect cell apoptosis. Moreover, p38 mitogen-activated protein kinase (MAPK) phosphorylation and C/EBP homologous protein (CHOP) expression were detected with Western blot. The results showed that Cav-1 expression was markedly increased at early stage of TG treatment (P < 0.05) and then decreased with prolonged or high dose TG treatments. The increasing of Cav-1 expression induced by TG in RAW264.7 cells was abolished under inhibition of caveolae by filipin(III) (P < 0.05). The effect of TG on apoptosis of RAW264.7 cells was further augmented after pretreatment with filipin(III) (P < 0.05). Western blotting showed that MAPK phosphorylation induced by TG was inhibited by filipin(III) in RAW264.7 cells (P < 0.05), whereas CHOP remained unchanged (P > 0.05). These results suggest that Cav-1 may play a critical role in suppressing ER stress-induced macrophages apoptosis in vitro, and one of the mechanisms may be correlated with the activation of p38 MAPK prosurvival pathway.
Animals ; Apoptosis ; drug effects ; Caveolin 1 ; genetics ; metabolism ; Cell Line ; Endoplasmic Reticulum Stress ; physiology ; Filipin ; pharmacology ; MAP Kinase Signaling System ; Macrophages ; cytology ; drug effects ; Mice ; Thapsigargin ; pharmacology ; Transcription Factor CHOP ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

To investigate the role and mechanism of ceramide (Cer) regulation in alcohol-induced neuronal proliferation and the newborn neurons formation, we used sphingomyelin synthase 2 (predominant enzyme of Cer metabolism) knockout (SMS2(-/-)) and wild type (WT) female mice to establish the model of prenatal alcohol exposure. In 24 h after being given birth (postnatal day 0, P0), the offspring of model mice received blood sphingomyelin (SM) measurement with enzymatic method. On P0, P7, P14 and P30, the proliferation of granule cells in the dentate gyrus and newborn neurons were investigated with immunofluorescent labeling. The expression of protein kinase Cα (PKCα) in the hippocampus was tested with Western blot analysis. The results showed that the SM level of blood in SMS2(-/-) pups was significantly lower than that in WT pups. No matter in SMS2(-/-) or WT mice, the prenatal alcohol exposure down-regulated the SM levels in pups with dose-dependency. In both SMS2(-/-) and WT pups, the number of proliferative neurons and newborn neurons in the dentate gyrus gradually decreased with the growing age. Compared with the WT pups, SMS2(-/-) pups showed significantly more proliferative neurons and newborn neurons in the dentate gyrus. Notably, prenatal alcohol exposure dose-dependently increased proliferative neurons and newborn neurons in the dentate gyrus in both WT and SMS2(-/-) pups. The hippocampal expression of PKCα protein in SMS2(-/-) mice was lower than that in WT mice, and prenatal alcohol exposure could up-regulate the PKCα protein expression in both WT and SMS2(-/-) mice with dose dependency. These results suggest that alcohol exposure during pregnancy can induce the compensatory neural cell proliferation and the production of newborn neurons in offspring, and the Cer-ceramide-1-phosphate (C1P) pathway is involved in alcohol-induced neural cell proliferation. The activation of PKCα may be a key step to start the Cer-C1P pathway and up-regulate the alcohol-induced neural cell proliferation and the newborn neurons formation.
Animals ; Animals, Newborn ; Cell Proliferation ; drug effects ; Cells, Cultured ; Ceramides ; metabolism ; Dentate Gyrus ; cytology ; Ethanol ; toxicity ; Female ; Mice ; Mice, Knockout ; Neurons ; cytology ; Pregnancy ; Prenatal Exposure Delayed Effects ; physiopathology ; Protein Kinase C-alpha ; metabolism ; Signal Transduction ; Transferases (Other Substituted Phosphate Groups) ; genetics

Objective The objective of this study was to evaluate the correlation between maximum standard uptake values( SUVmax ) of 18 F-FDG PET/CT and BRAF mutation in patients with papillary thyroid car-cinoma(PTC).Methods Atotalof51patients(meanageof49.3±12.9years)whounderwent18F-FDGPET/CT imaging and biopsy before thyroidectomy were retrospectively analyzed. Based on the pathological results, there were 48 patients with PTC and 3 patients with follicular thyroid carcinoma(FTC). The SUVmax of thyroid nodule was measured by semi-quantitative analysis. The correlation between clinical data( gender,age,tumor size and thyroglobulin concentration) and SUVmax was also analyzed. The difference of SUVmax between the two groups was analyzed by comparing the BRAF V600E mutation group and the non-mutation group. Results In patients with PTC,27 patients had BRAF V600E mutations and 11 patients had no tumor mutation. The SUVmax of BRAF V600E mutation group was significantly higher than that in the non-mutated group(5. 5 ± 3. 9 vs. 2. 2 ± 1. 2,P=0. 002). The SUVmax of patients with tumor diameter≥1 cm was significantly higher than that in patients with tumor <1 cm(P<0. 05). The SUVmax of patients with elevated thyroglobulin concentration was higher than that in normal patients(P<0. 05). No BRAF V600E mutation was observed in the FTC group. Conclusion The BRAF V600E mutant gene has a high SUVmax value in patients with PTC. There were significant difference in SUVmax among different tumor size and serum thyroglobulin concentration.

BACKGROUND:Schwann cells are important celllines in the process of repairing peripheral nerve injury, and human amniotic homogenate supernatant is shown to secrete a variety of cytokines, which could promote the proliferation of Schwann cells.
OBJECTIVE:To investigate the effect of different concentrations of human amniotic homogenate supernatant on the proliferation of rat Schwann cell96.
METHODS:Schwann cell96 was cultured with high-glucose DMEM containing 20%fetal bovine serum, and the second generation of Schwann cell96 was applied for experiments. The cultured cells were divided into five groups according to different volume fractions of human amniotic homogenate supernatant (0%, 10%, 15%, 20%, 25%) in the medium.
RESULTS AND CONCLUSION:The total protein concentration of human amniotic homogenate supernatant was 675μg/mL, in which the concentration of epidermal growth factor, basic fibroblast growth factor and vascular endothelial growth factor were respectively (470.625±2.546), (4.121±0.026) and (0.172±0.002) ng/L. At 1-7 days, the cellproliferation rate of the 10%and 15%concentration groups was greater than that in 20%and 25%concentration groups (P<0.05);10%and 15%concentrations promoted cellproliferation, while 20%and 25%concentrations inhibited cellproliferation. There were no significant difference in the viability of Schwann cell96 between the control group and the experimental group (P>0.05). Low concentrations (10%, 15%) of human amniotic homogenate supernatant promote the proliferation of Schwann cell96, while high concentrations (20%, 25%) of human amniotic homogenate supernatant inhibit cellproliferation.

Objective To explore standardized continuing vascular ultrasound training in China.Methods Data of current ultrasound practitioner education levels in the secondary and tertiary hospitals were collected.Under the direction of the Chinese Ultrasound Doctor Association (CUDA),Vascular Ultrasound Guidelines were introduced.The CUDA Steering Committee for Standardized Vascular Ultrasound Training and Vascular Ultrasound Training Centers were established.Results The questionnaires were received from 182 ultrasound departments of 57 secondary hospitals and 125 tertiary hospitals.The proportion of ultrasound practitioners who obtained a university degree was significantly higher in the ultrasound departments of tertiary hospitals than that in the secondary hospitals (75.18% vs 52.63%).The proportion of ultrasound practitioners who only received short-term ultrasound training and did not have tertiary education was significantly higher in the ultrasound departments of secondary hospitals than that in the tertiary hospitals (8.31% vs 1.03%).The directors of ultrasound departments in both secondary and tertiary hospitals believed that ultrasound practitioners should at least have a university diploma and preferably have a university degree.The CUDA and the American Registry for Diagnostic Medical Sonography (ARDMS) co-hosted two standardized vascular ultrasound training seminars and the first Registered Physician in Vascular Interpretation (RPVI) credential examination in Beijing.A well-known vascular ultrasound reference book,Introduction to Vascular Ultrasound was translated from English into Chinese and published in September 2008.The CUDA Vascular Ultrasound Guidelines were drafted by 12 vascular ultrasound experts from China,U.S.and Australia,which were widely discussed and passed by the CUDA Standing Committee.The guidelines have been published in the Chinese Journal of Ultrasonography and on the website of CUDA (www.cuda.org.cn).The CUDA has signed a publishing contract with a medical press to publish the guideline booklet.The CUDA Steering Committee for the Standardized Vascular Ultrasound Training was established which includes 24 domestic and foreign experts.The CUDA Standardized Vascular Ultrasound Training Centers were awarded to 12 hospitals,which were selected,trained and assessed by the CUDA from 35 candidates.Each center is expected to undertake one to two standardized vascular ultrasound training courses each year.Conclusion Continuing education of vascular ultrasound introduced by CUDA has been accepted positively by a portion of Chinese doctors with ultrasound major.