Objective:To analyze, at cellular level, whether the mouse B16-F1 melanoma cells with OAZI-1 overexpression could activate antigen-presenting cells and promote the phagocytotic and antigen-presenting efficiencies of mouse peritoneal macrophage and bone marrow derived DC on tumor cells. Methods:The plasmid pcDNA3. 1(+)/OAZI-1 was transfected into B16-F1 cells by Li-pofectamine2000 reagent. The positive clones with OAZI-1 overexpression ( B16/OAZI-1 ) were identified by Western blot assay and RT-PCR. Macrophages from abdominal cavity and DC from bone marrow were collected from BALB/c mouse. The B16-F1 cells transfected with the pcDNA3. 1(+) (B16/3. 1) were used as the control cells in this experiment. B16-F1 cells and macrophages were co-cultured for 4 h at a 1∶5 ratio and DC were co-cultured with B16-F1 cells at 1∶1 ratio for 4 h. And then the phagocytotic efficiencies were assayed by flow cytometry. DC were co-cultured with B16-F1 cells at 1∶1 ratio for 24 h and then the expression of mature DC surface marker molecules CD40,CD80,CD86 were determined by flow cytometry. The DC activated by the tumor cells were co-cultured with mouse spleen lymphocytes for 24 h, and then IFN-γ content in culture medium was analyzed by ELISA. Results: Phagocytotic assay showed that,compared to the control cells,the OAZI-1 overexpression in B16-F1 cells significantly enhanced the engulfment of B16-F1 cells by macrophages ( 24. 7% vs 53. 9% ) and DC ( 8. 2% vs 13. 8%) . When DC were co-incubated with OAZI-1 overexpressed B16-F1 for 24 h,the expression levels of CD40,CD80,CD86 on the DC surface,which were the molecular markers for matured DC,increased from 24. 2%,20. 8% and 16. 4% to 46. 8%,32. 5% and 36. 1% respectively. Co-culture of tumor-activated DC with the spleen lymphocytes resulted in an increased IFN-γcontent in the culture medium(32. 9 pg/ml vs 15. 1 pg/ml). Conclusion:The tumor cells with OAZI-1 overexpression can be engulfed more efficiently by macrophages and DC. And this process can induce the maturation and activation of DCs. Matured DC could induce T cell activation and then activate the anti-tumor immune response.

AIM:To investigate the gene expression profile of human lung squamous cell carcinoma in early stage.METHODS:The mRNA was extracted from cancer tissue and normal lung tissue.The mixed probes were labeled and hybridized with chip containing 480 carcinoma related genes,then analyzed by SuperArray Image software to study the gene expression patterns in lung squamous cell carcinoma.RESULTS:192 genes associated with lung squamous cell carcinogenesis were found,which were grouped into transporter,adhesion molecules,cytoskeleton proteins,transcription regulator,genes associated with metabolism and immune response according to functions.127 gens were positive to lung squamous cell carcinogenesis and 65 genes were negative to lung squamous cell carcinogenesis.CONCLUSION:The screen of gene expression profile of human lung squamous cell carcinoma provides valuable information for the research of molecular mechanism of the lung squamous cell carcinogenesis.

The combined application of statins that inhibit HMG-CoA reductase and fibrates that activate PPAR-α can produce a better lipid-lowering effect than the simple application, but with stronger adverse reactions at the same time. In the treatment of hyperlipidemia, the combined administration of TCMs and HMG-CoA reductase inhibitor in treating hyperlipidemia shows stable efficacy and less adverse reactions, and provides a new option for the combined application of drugs. In this article, the pharmacophore technology was used to search chemical components of TCMs, trace their source herbs, and determine the potential common TCMs that could activate PPAR-α. Because there is no hyperlipidemia-related medication reference in modern TCM classics, to ensure the high safety and efficacy of all selected TCMs, we selected TCMs that are proved to be combined with statins in the World Traditional/Natural Medicine Patent Database, analyzed corresponding drugs in pharmacophore results based on that, and finally obtained common TCMs that can be applied in PPAR-α and combined with statins. Specifically, the pharmacophore model was based on eight receptor-ligand complexes of PPAR-α. The Receptor-Ligand Pharmacophore Generation module in the DS program was used to build the model, optimize with the Screen Library module, and get the best sub-pharmacophore, which consisted of two hydrogen bond acceptor, three hydrophobic groups and 19 excluded volumes, with the identification effectiveness index value N of 2. 82 and the comprehensive evaluation index CAI value of 1. 84. The model was used to screen the TCMD database, hit 5,235 kinds of chemical components and 1 193 natural animals and plants, and finally determine 62 TCMs. Through patent retrieval, we found 38 TCMs; After comparing with the virtual screening results, we finally got seven TCMs.
Acyl Coenzyme A ; metabolism ; Animals ; Databases, Factual ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Hydrophobic and Hydrophilic Interactions ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; chemistry ; pharmacology ; Lipid Metabolism ; drug effects ; Lipids ; blood ; Medicine, Chinese Traditional ; Models, Molecular ; Quantitative Structure-Activity Relationship ; Technology

Objective To observe effect of Compound Ruikangxin Capsula(CRC) on anxiety-like symptoms of morphine-withdrawal rats and investigate its mechanism.Methods The elevating doses were applied to validating anxiety-like behavior in SD male rats with sc morphine for 10 d.Treatments with ig administration of CRC(100,200,and 300 mg/kg) and buspirone(15 mg/kg) were performed during 1—3 d morphine withdrawal(twice every day).The plus elevated maze was applied to validating anxiety-like symptom in rats.The expression levels of synaptophysin were detected on hippocampus in groups by immunohistochemitry 72 h after sc morphine.Results As compared with the control group,animals treated with CRC(200 and 300 mg/kg) and buspirone had higher ratio of entry into open arm (P

Objective To investigate the epidemiology of viral infection in elderly patients to contract acute exacerbations of chronic obstructive pulmonary disease (AECOPD)in Minhang district of Shanghai from 2010 to 2012,and to study the relationships between viral infection and clinical features.Methods The elderly patients (age >70 year old)with AECOPD admitted from September 2010 to November 2012 were enrolled for study.The patients who couldn't complete lung function test were excluded.The pharyngeal swabs (PS)were taken from each patient within the first 24 h after admission.Nine respiratory viruses and their subtypes from pharyngeal swabs were detected by the nested multiplex polymerase chain reaction (PCR)method,including influenza virus A (FluA),2009 influenza A (H1N1 )virus (09FluH1 ), influenza virus B (FluB),respiratory syncytial virus A (RSVA)and B (RSVB),human coronavirus-229E (hCOV-229E),human coronavirus-NL63 (hCOV-NL63 ),human coronavirus-OC43 (hCOV-OC43 ), human coronavirus-HKU1 (hCOV-HKU1),human parainfluenza virus 1-4 (hPIV1-4),human adenovirus (hAdV),human boca virus (hBoV),human metapneumo-virus (hMPV)and human rhinovirus (hRV). According to the PCR results,all patients were divided into positive viral infection group and negative viral infection group.The relationships between viral infection and clinical features were analyzed.Results Sixty patients were eligible for study.Of them,14 patients were found to be positive for virus infection including a triple infected patient (FluB,hRV and hROV)and 46 patients were negative for virus infection.The viral pathogens detected in the positive viral group were:9 cases of hRV (15.00%),2 cases of hPIV (3.33%),2 cases of hCOV (3.33%),2 cases of FluB (3.33%)and 1 cases of RSV (1.67%).The mortality in the positive viral group was higher than that in the negative viral group.However,the other clinical characteristics between the two groups had no significant differences. Conclusions Human rhinovirus was the most common viral pathogen in elderly patients with AECOPD.Viral infection might be associated with the prognosis.However,the patients with viral infection are lack of specific clinical characteristics,therefore,the prompt diagnosis before careful study would be difficult.

OBJECTIVETo systematically evaluate acupuncture as a treatment for male infertility.

METHODSWe searched Chi na Biology Medical Database (CBM), Wan Fang Medical Information System, China National Knowledge Infrastructure (CNKI), VIP Information Resource System (VIP), and PubMed for published literature on acupuncture as a treatment for male infertility on May 1 2014. Based on the Standards for Reporting Interventions in Clinical Trials of Acupuncture (STRICTA), we evaluated the quality of the reports, conducted meta-analysis on the identified studies via RevMan5.2, and assessed the quality of the evidence in the literature by Grading of Recommendations Assessment, Development, and Evaluation (GRADE).

RESULTSA total of 12 studies involving 2,177 patients were included, the quality of which was evaluated as mediocre. With regard to the cure rate, acupuncture was comparable to traditional Chinese medicine (TCM) (P > 0.05) but better than Western medicine (RR = 4.00, 95% CI 1.63 to 9.82, P < 0.01) while acupuncture + TCM was better than either TCM (RR = 1.77, 95% CI 1.20 to 2.60, P < 0.01) or Western medicine used alone (RR = 2.73, 95% CI 1.51 to 4.93, P < 0.01), and acupuncture + Western medicine was better than Western medicine alone (RR = 1.88, 95% CI 1.17 to 3.02, P = 0.01). The combined use of acupuncture, ear pressure, TCM, and Western medicine showed a higher cure rate than the combination of TCM and Western medicine (RR = 3.45, 95% CI 2.90 to 4.11, P < 0.01). In therapeutic effectiveness, acupuncture was comparable to TCM (P > 0.05) but superior to Western medicine (RR = 1.41, 95% CI 1.12 to 1.71, P < 0.01), acupuncture + TCM was superior to either TCM (RR = 1.14, 95% CI 1.05 to 1.23, P < 0.01) or Western medicine alone (RR = 1.43, 95% CI 1.22 to 1.67, P < 0.01), and acupuncture + Western medicine was superior to Western medicine alone (RR = 1.25, 95% CI 1.05 to 1.49, P = 0.01). In improving sperm concentration, acupuncture was as effective as TCM (P > 0.05) and sham acupuncture (P > 0.05) but outdid Western medicine (RR = 27.00, 95% CI 24.27 to 29.73, P < 0.01) and acupuncture + TCM outdid either TCM (RR = 14.65, 95% CI 7.58 to 21.72, P < 0.01) or Western medicine alone (RR = 1.04, 95% CI--1.43 to 3.51, P > 0.05). In improving grade a sperm, acupuncture exhibited a similar effect to TCM (P > 0.05) and sham acupuncture (P > 0.05), and acupuncture + TCM was more effective than TCM alone (RR = 7.78, 95% CI 3.51 to 12.23, P < 0.01) but equally effective as Western medicine (P > 0.05). In elevating the level of grade a + b sperm, acupuncture + TCM excelled either TCM (RR = 11.00, 95%, CI 3.17 to 18.82, P < 0.01) or Western medicine alone (RR = 12.22, 95% CI 6.87 to 17.57, P < 0. 01), while acupuncture produced a comparable effect with sham acupuncture (P > 0.05). As for the quality of the included studies, only 3 conclusions of the 23 meta-analyses were assessed to be of average quality, while the others of poor or extremely poor quality. Therefore, the recommendation grade of the conclusions was low.

CONCLUSIONFor the treatment of male infertility, acupuncture is reported to be equally effective as TMC and more effective than Western medicine, and its effectiveness is enhanced when applied in combination with either TCM or Western medicine. Acupuncture is distinctively efficacious in improving sperm quality. Nevertheless, the overall quality of the included studies is low.

Cholesterol ester transfer protein (CETP) is a key regulator of high density lipoprotein (HDL). Owing to its important role in the reverse of cholesterol transport, CETP has become a hotspot target in modulating lipid drug design. In this paper, structure based pharmacophore (SBP) models for CETP inhibitors were built based on the protein structure 4F2A from Protein Database (PDB). The best pharmacophore contained six hydrophobic features, one hydrogen bond acceptor feature and nine excluded volume features, with the N and CAI value was 3.33 and 2.31 respectively. Then the model was used to search the traditional Chinese medicine database (TCMD) and 629 compounds originated from 315 TCM herbs were obtained. Molecular docking was also used to validate SBP by analyzing the critical amino acid residue and the interaction between potential active compounds and receptor. In this study, several TCM herbs, like Lycii Frutus and Schisandrae chinensis fructus, which contained more optimal SBP based screening results, have been reported hypolipidemic effect, and need to be studied deeply in a more focused research on herbal active constituents. Therefore, this study could provide reliable fundamental data for exploring the action mechanisms of TCM, and be applicable to identify lead candidates, which can be utilized as starting scaffolds for natural CETP inhibitors.
Cholesterol Ester Transfer Proteins ; antagonists & inhibitors ; Drug Evaluation, Preclinical ; methods ; Medicine, Chinese Traditional ; Molecular Docking Simulation

Objective: To establish a method for isolation and purification of fetal membrane derived adherent cells (FMDACs) , and investigate their biological characteristics. Method: FMDACs were isolated with trypsin inducing and cultured in vitro. FMDACs were induced to differentiate into osteoblasts and adipocytes. FACS and immunocytochemistry technique were used to examine the cell surface antigen. The genetic stability was verified by karyotype analysis. Results: FMDACs were successfully isolated and expanded in vitro. They had strong proliferative ability. FMDACs were positive for CD44 and CD29, but negative for CD34, CD14 and CD45. FMDACs were differentiated into osteoblasts and adipocytes after inducement. The karyotype was stable in the sixth-passaged FMDACs and the tumorigenicity was not found. Conclusion; FMDACs have the possibility of multipotent stem cells, which have strong capacities of self-renewal and multidirectional differentiation. The genetic background of FMDACs is stable. FMDACs may be used as a kind of novel seed cells for tissue engineering.

Exosomes are membrane vesicles secreted by cells into the extracellular environment. Exosomes are a new cell component in the treatment of spinal cord injury. Exosomes are secreted by mesenchymal stem cells have anti-apoptotic, anti-inflammatory and pro-angiogenic effects in the treatment of spinal cord injury, which play a key role in nerve cell-cell communication and can delivery exogenous genetic material. Exosomes can penetrate blood-brain barrier and are more stable than their parent cells, reducing the safety risks in the administration of viable cells.