OBJECTIVEEffects of ginsenoside Rb1, Rg1 and Re on neurotrophic factor signal transduction pathway using liposome-mediated transfection of eukaryotic cells approach.

METHODThe injury model was established by treating SH-SY5Y cells with 0.6 mmol x L(-1) of corticosterone (CORT) by 24 h. SH-SY5Y cell were pretreated with CORT for 30 min followed by co-treated with 120,60 and 20 micromol x L(-1) of Rb1, 120, 80 and 40 micromol x L(-1) of Rg1 and 120, 80 and 40 micromol x L(-1) of Re for 24 h. Cells viability was determined by Cell Counting Kit (CCK) assay. CREB expressing Luciferase reporter gene was constructed and transfected with plasmid containing hRaf, hcAMP, hAkt, hCaMK gene into human embryonic kidney (HEK293) cells using liposornal transfection reagent lipofection 2000. The expression of CREB before and after it addion of Rb1, Rg1 and Re was examined by Luc assay system and Western blotting.

RESULTCompared with normal control group, CORT significantly decreased the viability of SH-SY5Y cells to 67.21% (P < 0.01). CCK results show that Rb1 (60 micromol x L(-1)), Rg1 (80 micromol x L(-1)) and Re (80 micromol x L(-1)) on SH-SY5Y cells have significant protective effect (P < 0.01). Lucassay and Western blotting results show that the gene and protein levels of CREB increased significantly through the pathway of Raf and Akt with Rb1 and Rg1 (P < 0.01), Re can increase significantly the gene and protein levels of CREB through the pathway of Raf and CaMK II.

CONCLUSIONRb1, Rg1 and Re protects SH-SY5Y cells from CORT-induced damage and the neuroprotective mechanism may be associated with the Raf-CREB, Akt-CREB and CaMK II -CREB pathways.

Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Cell Line ; Cell Survival ; drug effects ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Genes, Reporter ; Ginsenosides ; pharmacology ; Humans ; Panax ; chemistry ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; drug effects ; raf Kinases ; genetics ; metabolism

Dingzhi Xiaowan is a widely used traditional Chinese medicine in treating depression, which is a similar formula of Kaixinsan. In this research, a rapid ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS(E)) method was established to analyze the metabolites of Dingzhi Xiaowan in depressive model rat plasma, bile, urine and feces. After we established Chronic unpredictable mild stress (CUMS) model rats and orally administrated Dingzhi Xiaowan, rat plasma, bile, urine and feces samples were collected and prepared. Using Waters Cortects UPLC C18 column (2.1 mm x 50 mm, 1.6 μm), acetonitrile-0.1% formic acid mobile phase gradient, these samples were analyzed and 33 metabolites of nine bioactive compounds were detected and tentatively identified by Metabolynx. Among the 33 metabolites, three metabolites were identified from plasma sample, three came from bile sample, and 27 metabolites were identified from urine and feces samples. This approach provided a rapid method for characterizing the metabolites of Dingzhi Xiaowan and gave the truly active structures and the action mechanism of their antidepressant effects.
Animals ; Bile ; metabolism ; Chromatography, High Pressure Liquid ; Depression ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; metabolism ; Feces ; chemistry ; Male ; Mass Spectrometry ; Medicine, Chinese Traditional ; Plant Extracts ; metabolism ; Rats ; Rats, Sprague-Dawley

The efficacy of Chinese herbal formula in treating depression has been proved in many studies. In this study, six different Kaixin San formulas were compared to investigate their effects on central monoamine neurotransmitters of chronic stress rats and against depression based on their different components in plasma, in order to discuss the efficacy-comparability relationship and the possible efficacy mechanism. The classic isolation method and the chronic unpredictable mild stress (CUMS) depression model were combined to investigate the changes in contents in hippocampus and monoamine neurotransmitters (NE, DA, 5-HT) and the components of some formulas in plasma with HPLC and UPLC-Q-TOF-MSE methods. As a result, Dingzhi Xiaowan recorded in Essential Recipes for Emergent Use Worth A Thousand significantly increased the behavioral scores, NE and 5-HT contents in hippocampus and NE, DA and 5-HT contents in cortex, with the best anti-depressant effect. Dingzhi Xiaowan recorded in Complete Records of Ancient and Modern Medical Works showed a notable increase in sucrose preference and open field score in model rats, NE content in hippocampus and NE, DA and 5-HT contents in cortex, with a certain anti anti-depressant effect. Kaixin San recorded in Ishinpo showed remarkable rise in weight of model rats. NE content in hippocampus and DA content in cortex. Puxin Decoction recorded in A Supplement to Recipes Worth A Thousand Gold showed 5-HT content in hippocampus and DA content in cortex. Kaixin San recorded in Yimenfang only showed DA content in cortex. Kaixin Wan recorded in Essential Recipes for Emergent Use Worth A Thousand did not mention the antidepressant effect. According to the results, the formulas' different anti-depressant effects may be related to the different plasma components.
Animals ; Behavior, Animal ; drug effects ; Biogenic Monoamines ; analysis ; Brain Chemistry ; drug effects ; Chronic Disease ; Drugs, Chinese Herbal ; pharmacology ; Male ; Medicine, Chinese Traditional ; Neurotransmitter Agents ; analysis ; Norepinephrine ; analysis ; Rats ; Rats, Sprague-Dawley ; Serotonin ; analysis ; Stress, Psychological ; metabolism

BACKGROUNDTransradial coronary intervention has been widely used because of its effects in lowering the incidence of complications in vascular access site and improving patient satisfaction compared to the femoral approach. This study aimed to investigate the safety and feasibility of transradial approach for primary percutaneous coronary intervention (PCI) in elderly patients with acute myocardial infarction (AMI).

METHODSA total of 103 consecutive elderly patients (age = 65 years) who were diagnosed as having AMI were indicated for PCI. Among them, 57 patients received primary PCI via the transradial approach (transradial intervention, TRI group), and 46 underwent primary PCI via the transfemoral approach (transfemoral intervention, TFI group). The success rate of puncture, puncture time, cannulation time, reperfusion time, the total time for PCI, the success rate of PCI, the use rates of temporary pacemaker and intra-aortic balloon pump (IABP), and the total length of hospital stay of the patients in the two groups were compared. After the procedure, vascular access site complications and major adverse cardiovascular events (MACE) in the two groups in one month were observed.

RESULTSThe success rates of puncture (98.2% vs 100.0%) and PCI (96.5% vs 95.7%) for the patients in the TRI and TFI groups were not statistically significant (P > 0.05). The puncture time ((2.4 +/- 1.1) vs (2.0 +/- 0.9) minutes), cannulation time ((2.7 +/- 0.5) vs (2.6 +/- 0.5) minutes), reperfusion time ((16.2 +/- 4.5) vs (15.4 +/- 3.6) minutes), total time of the procedure ((44.1 +/- 6.8) vs (41.2 +/- 5.7) minutes), use rates of temporary pacemaker (1.8% vs 2.2%) and IABP (0 vs 2.2%) in the two groups were not statistically significant (P > 0.05), but the hospital stay of the TFI group was longer than that of the TRI group ((10.1 +/- 4.6) vs (7.2 +/- 2.6) days, P < 0.01). A radial occlusion was observed in the TRI group, but no ischemic syndrome in hand. In the TFI group, 4 patients had hematosis, 1 had pseudoaneurysm, and 1 had major bleeding. Statistical significance in vascular access site complications was seen in the two groups (1.8 % vs 13.1%, P < 0.05). Three patients died in the two groups respectively in one month, and there was no statistical significance in MACE in the two groups (5.3% vs 6.5%, P > 0.05).

CONCLUSIONThe transradial approach for primary PCI is safe and feasible for elderly patients with AMI.

Aged ; Angioplasty, Balloon, Coronary ; adverse effects ; methods ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; therapy ; Radial Artery

OBJECTIVETo investigate the role of Smac in cisplatin-induced apoptosis of non-small lung cancer cells in vitro.

METHODSNon-small cell lung cancer A549 cells were incubated in the presence of cisplatin at different concentrations, and the cell proliferation status was observed using MTT assay. Flow cytometry was used for evaluation of the apoptosis of the incubated cells, and the expressions of Smac mRNA and protein were detected by RT-PCR and Western blotting, respectively.

RESULTSCisplatin inhibited the proliferation and induced apoptosis of A549 cells both in a concentration-dependent manner. Cisplatin also increased the expression of Smac at both the mRNA and protein levels, which was also concentration-dependent.

CONCLUSIONIncreased Smac expression may play a critical role in cisplatin-induced apoptosis of the non-small cell lung cancer cells in vitro.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Inhibitor of Apoptosis Proteins ; biosynthesis ; genetics ; Intracellular Signaling Peptides and Proteins ; genetics ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Mitochondrial Proteins ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To discuss the common complications of large acoustic neuromas performed surgery via suboccipital retrosigmoid approach under neurophysiological monitoring, and the prevention of postsurgical complications. Methods One hundred and sixteen patients with large acoustic neuromas, admitted to our hospital from May 2006 to April 2010, were performed surgery via suboccipital retrosigmoid approach under neurophysiological monitoring; clinical data and follow-up data (3-12 months) of these patients were collected and analyzed retrospectively; the influence of tumor dimension on short- and long-term postoperative complications after the surgery were compared.Results Total tumor resection was achieved in 107 patients, subtotal resection in 7 patients, and partial resection in 2 patients. After the operation, the facial nerve was preserved anatomically in 101 patients (87.07%); and the functional valuation of facial nerve according to postoperative House-Brackman showed 89 patients (76.72%) in grade Ⅰ-Ⅱ, 13 patients (11.21%) in gradeⅢ-Ⅳ and 14 patients (12.07%) in grade Ⅴ-Ⅵ. Short-term postoperative complications included hoarse voice in 16 patients and herpes catarrhalis of injured side in 20 patients. Long-term postoperative complications included loss of hearing (n=83) and permanent facial paralysis (n=23). Patients with tumor dimension larger than 5 cm had a significantly higher incidence of short-term postoperative complications than those within 3-5 cm (P< 0.05). No patient died. Conclusion The keys to avoiding the complications are to master the clinical anatomy of the approach, accumulate surgical experiences, preoperatively study the individual imaging and clinical data, and use intraoperative nerve monitoring.

We report a case of in vitro fertilization and embryo transfer (IVF?ET) with oocyte donation in a woman with premature ovarian insufficiency (POI) complicated by systemic lupus erythematosus (SLE) during pregnancy. The patient had a diagnosis of POI 4 years earlier and 11 weeks after successful pregnancy by IVF?ET with oocyte donation in 2003, she presented with facial edema, and further examinations confirmed the diagnosis of lupus nephritis. She received treatment with prednisone to control the activity of SLE and aspirin and low?molecular?weight heparin to improve placental blood flow with close monitoring of gravida and fetus throughout pregnancy. The condition of the patient remained unstable during pregnancy, and liver damage and placental circulation disorder occurred in late gestational weeks with suspected intrauterine growth retardation (IUGR) of the fetus. For maternal and fetal safety, the patient received elective caesarean section and delivered a premature boy at 31 weeks of gestation. She subsequently received further medications for SLE and showed good recovery of the immunological parameters and absence of SLE symptoms during the follow?up for 14 years, indicating a clinical cure of SLE. Her son shows normal growth and development. Based on the experience with this case and literature review, we believe that immunological factor is an important cause of POI and thus recommend full immunological examinations in cases of idiopathic POI.

We investigated epidemiological characteristic of scrub typhus in Yongshan County,Yunnan Province,China. The serum samples were collected from the patients with fever for detecting the antibody against Orientia tsutsugamsushi (Ot) by colloidal gold immunoassay assay.Rat traps were used to capture rodents.The spleen tissues of the captured rodents were detected by nested-polymerase chain reaction for rickettsia groEL segment.The groEL segments were sequenced and analyzed the homology with the other known sequences.Thirty-four scrub typhus cases were found in Yongshan County,Yunnan Prov-ince from May 2015 to October 2015.Among them,21 cases were confirmed by laboratory tests and 13 cases were clinical di-agnosis diseases.Of these patients,32.35% of the cases occurred in June.The 32.35% were in the group of the 40-49 year-old,and 79.41% were farmers,94.12% exhibited eschar or skin ulcer(31.25% were observed in groin of these cases),and rash developed in 50%.In 39 spleen tissue samples of Rattus flavipectus,9 samples showed positive for groEL gene Ot,but gro-EL gene of Typhus group rickettsia and spotted fever group rickettsia were negative.Sequence analysis showed that YSP30 was closely related to some Saitama related strains of Ot,such as HSB1,FAR1 and UAP4,while the other 8 strains were closely related to some Karp related strains of Ot,such as UT213,UT221 and SH205.It was confirmed that the Yongshan County was the natural foci of scrub typhus by the serological and molecular biological detections.There are Karp and Saitama genotype related Ots in the natural foci.

Outer membrane vesicles (OMVs) are nanoscale vesicles secreted by Gram-negative bacteria. As a unique bacterial secretion, OMV secretion can help bacteria maintain the outer membrane stability or remove harmful substances. Studies have shown that local separation of outer membrane and peptidoglycan layers led by abnormalities in outer membrane protein function, abnormal structure or excessive accumulation of LPS, and erroneous accumulation of phospholipids in the outer leaflet, which can all lead to bacterial outer membrane protrusion and eventually bud formation of OMVs. Since OMVs are mainly composed of bacterial outer membrane and periplasmic components, the pathogen associated molecular patterns (PAMPs) on their surface can trigger strong immune responses. For example, OMVs can recruit and activate neutrophils, polarize macrophages to secrete large amounts of inflammatory factors. More importantly, OMVs can act as adjuvants to induce dendritic cell (DC) maturation to enhance adaptive immune response in the body. At the same time, OMVs are derived from bacteria, which make it easy to modify. The methods by genetic engineering and others can improve their tumor targeting, give them new functions, or reduce their immunotoxicity, which is conducive to their application in tumor therapy. OMVs not only induce apoptosis or pyroptosis of tumor cells, but also regulate the host immune system, which makes OMVs themselves have a certain killing effect on tumors. In addition, the tendency of neutrophils to inflammatory tumor sites and the formation of neutrophil extracellular traps enable OMVs to target tumor sites, and the suitable size and the characteristic that they are easily taken up by DCs give OMVs a certain lymphatic targeting ability. Therefore, OMVs are often employed as excellent drug or vaccine carriers in tumor therapy. This review mainly discusses the biological mechanism of OMVs, the regulatory effects of OMVs on immune cells, the functional modification strategies of OMVs, and their research progress in tumor therapy.

Objective:To explore the possible mechanism of Huangqintang in treating ulcerative colitis （UC）. Method:The animal model of UC was induced by dextran sodium sulfate （DSS）．The experimental animals were divided into control group， model group，Huangqintang low dose （4.55 g·kg^-1）， medium dose （9.1 g·kg^-1）， and high dose（18.2 g·kg^-1） groups. Intragastric administration was also given in the modeling process for 7 consecutive days. At the end of the 8th day， colon tissues were collected to measure colon length and mass， and calculate the colon mass index. Pathological changes were observed by hematoxylin-eosin （HE） staining. Serum iron content， superoxide dismutase （SOD）， glutathione （GSH）， catalase （CAT） and myeloperoxidase （MPO） were determined by biochemical assay. Western blot was used to detect the protein expression of glutathione peroxidase 4 （GSH-Px4）， long-chain acyl-CoA synthetase 4 （ACSL4） and ferritin heavy chain 1（FTH1）. The mRNA expression levels of tumor trotein 53 （P53） and solute carrier family 7 member 11 （SLC7A11） in colon tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:The experimental studies showed that compared with normal group， serum MPO and iron content， ACSL4 protein level and relative P53 mRNA expression in the model group significantly increased （P<0.05）， while serum SOD， CAT， GSH content， GSH-Px4， FTH1 relative protein expression level and relative SLC7A11 mRNA expression in the model group significantly decreased （P<0.01）. Compared with model group， serum MPO and iron content， ACSL4 protein level and relative P53 mRNA expression significantly decreased （P<0.05）， while serum SOD， CAT， GSH content， GSH-Px4， FTH1 relative protein expression level and relative SLC7A11 mRNA expression significantly increased （P<0.05） after the intervention of Huangqintang， and the effect was most significant in the high-dose group （P<0.05）. The results of general condition， colon length， colon mass index and HE staining showed that Huangqintang could relieve clinical symptoms and histopathological changes in UC mice. Conclusion:These results indicated that Huangqintang had therapeutic effect on ulcerative colitis mice， and its mechanism might be related to inhibiting the oxidative stress and ferroptosis.