AIM: To observe the changes of the number of NKT cells in spleens and livers of induced model of experimental autoimmune encephalomyelitis (EAE), and to study the role NKT cells play in the immunoregulation of EAE. METHODS: C57BL/6 mice were immunized with MOG<,35-55> peptide and received clinical evaluation daily. The mice were sacrificed at the fastigium and the splenic and hepatic lymphocytes were isolated. The changes of NKT cells in normal and EAE C57BL/6 mice were detected by flow cytometry. RESULTS: The percent of NKT cells in lymphocytes of different organs of EAE model were greater decreased than in that of normal mice. The percent of NKT cells in splenic lymphocytes of normal mice was 2.22± 0.14, while that in EAE mice was 1.94±0.07 (P < 0.05). The percent of NKI cells in hepatic lymphocytes of normal mice was 5.52±2.17, while that in EAE mice was 2.67± 1.41 (P < 0.05). CONCLUSION: The proliferation of splenic and hepatic NKT cells in C57BL/6 mice are inhibited in EAE model, which may indicate that the immune function conducted by NKT cell is down regulated in EAE mice.

Objective To analyze the methods and effects for treating unstable posterior pelvic ring fracture combined with sacral nerve injury and further identify the relationships among the diagnostic methods,surgical approaches and clinical outcomes.Methods A total of 38 patients with posterior pelvic ring fracture combined with sacral plexus injury treated from January 2000 to January 2010 were enrolled in the study.There were 20 males and 18 females at an average age of 35 years (range,10 to 59 years).The causes of fractures included traffic injury in 20 patients,fall injury in 12,weighty object impingement injury in five,and stabbing injury in one.Classification of posterior pelvic ring fractures included fracture and dislocation of sacroiliac joints in eight patients,fracture of ilium wing in two and sacrum fracture in 28.According to the Denis typing of sacrum fractures,there was one patient with type Ⅰ fracture,14 with type Ⅱ fracture and 13 with type Ⅲ fracture.All 38 patients presented the decrease or loss of skin sensation around the lower extremities,perineal region and crissum.Simultaneously,30 patients suffered motor dysfunction of the lower extremities,while 20 patients had bladder and anus sphincter dysfunction or sexual disorder.Thirteen patients were suspected of sacral plexus avulsion and four of them were confirmed by myelography or MRI examination.All patients had at least one associated injury.The average ISS was 21.9 points ( range,9 to 47 points).Therapeutic methods were fracture reduction and fixation in the absence of nerve decompression for eight patients and nerve decompression for 30 patients including 26 patients being also managed by fracture reduction and fixation.Operation time ranged from 6 days to 6 months.The clinical outcomes were evaluated according to the British Medical Research Council (BMRC) evaluation criteria of sensation and movement function.Results Thirty-four patients were followed up for average 4.9 years ( range,1 to 10 years),during which their pelvis obtained stable recovery.The neurological outcome was excellent in two patients,good in four and unchanged in two in the nondecompression group and was excellent in 16 patients,good in nine and unchanged in one in the decompression group,with the decompression group superior to the non-decompression group ( P ＜ O.05 ).Conclusions For unstable posterior pelvic ring fracture combined with sacral nerve injury,nerve decompression and release combined with internal fixation can better improve the sacral nerve function and obtain good pelvic ring stability and is worth of clinical application.

OBJECTIVEThis study investigates construction of cardiac muscle cell-porous collagen scaffold complex in a bioreactor so as to unveil the possibility of generating 3-dimensional cardiac muscle tissue under the environment that mimics microgravity in vitro.

METHODS1-2-day old neonatal rat cardiac muscle cells were isolated by sequential digestion and pre-plating methods, then seeded onto porous collagen scaffold. The cell-collagen complex was transferred into rotary cell culture system (RCCS) and incubated for 7 days. Cells cultured in 75 ml flasks and constructs cultures on plates served as control. Morphological changes of the cells were observed by light microscope and metabolic rate was recorded. Ultrastructure of the cells growing in porous collagen was observed by transmission electron microscopy. Content of total DNA and protein in the newly-formed tissue were analyzed. H-E and anti-sarcomeric alpha-actin stains were performed in comparison with native cardiac muscle.

RESULTSThe isolated cardiac muscle cells adhered to the bottom of the flasks 24 hours after plating and began to beat spontaneously. When incubated for 7 days in RCCS, cell-collagen constructs of form a continuous outer tissue layer containing cells aligned with each other. The cell population in the interior of the construct was less in density than the outer part. Transmission electron microscopy demonstrated that subcellular elements characteristic of cardiac myocytes were in the outermost layer of constructs. A strongly positive stains of anti-sarcomeric alpha-actin suggested presence of cell population of differentiated cardiac myocytes in these constructs. Construct biomass was not significantly different from that in neonatal rat ventricle and approximately 40% of that in adult rat ventricles. Construsts in plates contained a few of cells which were less than those in RCCS. Metabolic activity of cells cultured in RCCS was higher than that in flasks and plates.

CONCLUSIONSDissociated cardiac muscle cells cultured on 3-dimensional scaffolds in RCCS under favorable conditions can form engineered constructs with structural and functional features resembling those of native cardiac tissue.

Animals ; Animals, Newborn ; Bioreactors ; Cell Division ; drug effects ; Cell Separation ; Cells, Cultured ; Collagen ; Culture Media ; Myocytes, Cardiac ; cytology ; Rats ; Tissue Engineering

With the kernel of efficacy, "Xiaohe Silian" was a pattern and method for new drug discovery which was constituted with "metabolism-efficacy, toxicity-efficacy, quality-efficacy and structure-efficacy". Its connotation was in keeping with traditional Chinese medicine (TCM) clinical pharmacy. This paper systematically summarized the research method of new drug discovery practice process for TCM. To avoid western drug like in TCM new drug discovery, we carried out combination analysis with TCM clinical pharmacy. The correlation analysis between basic elements of "Xiaohe Silian(n) and TCM clinical pharmacy was studied to guarantee this method could integrate closely with TCM clinic from all angles. Hence, this method aimed to provide a new method for TCM new drug discovery on the basis of TCM clinical pharmacy with insisting on holistic view of multicomponent study, kinetic view of metabolic process when the curative effect occurred and molecular material view of quality control and structure-activity exposition.
Drug Discovery ; methods ; Drug Therapy ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Humans ; Medicine, Chinese Traditional

Objective: To investigate the chemical constituents from the stems of Clausena emarginata. Methods: The compounds were isolated by macroporous resin, silica gel, ODS column chromatography, Sephadex LH-20, reversed-phase MPLC, and then purified by preparative HPLC. Their structures were determined by the analysis of ultraviolet spectrum, mass spectrum, and NMR spectrum. Neuroprotective activities of compounds 11 and 12 were initially investigated. Results: Sixteen compounds were isolated from the petroleum ether and acetone fractions of 95% ethanol extract of the stems of Cl. emarginata, and their structures were identified as 1H-Indole-3-carboxaldehyde (1), E-N-benzoiltiramine (2), dehydrodiconiferyl alcohol (3), tortoside A (4), zhebeiresinol (5), evofolin B (6), 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucopyranoside (7), echipuroside A (8), 3-methylcarbazole (9), murrayafoline A (10), clausine Z (11), indizoline (12), clausenaline B (13), mafaicheenamine A (14), dictamnine (15), and honokiol (16). Conclusion: Compounds 1-8 are isolated from the plants of genus Clausena L. for the first time, compounds 1-16 are isolated from this plant for the first time. Compounds 11 and 12 show the neuroprotective activity against rotenone induced PC12 cell damage.

Glucose-regulated protein 78 (GRP78) exerts critical influence on endoplasmic reticulum stress (ER stress),carcinogenesis and tumor progression.Cytochrome P450 (CYP450) enzyme is of great importance in the metabolism of drug and carcinogens.This article reviews recent studies on the interaction between GRP78 and CYP450 in order to provide evidence of novel strategies for cancer treatment.

OBJECTIVETo discuss the feasibility of estimating energy consumption of green laser in photoselective vaporization of prostate (PVP) based on the preoperative volume of prostate.

METHODSFrom January 2005 to January 2007, 260 patients with benign prostatic hyperplasia (BPH) had been treated with PVP. Preoperative prostatic volume and post-two-weeks-operative prostatic volume of each patient were measured by transrectal ultrasound (TRUS). Energy consumption and emission time of green laser were recorded during the operation. Then we calculated the amount of energy consumption needed in vaporizing one gramme prostatic tissue, evaluated the correlation of energy consumption and preoperative volume of prostate by means of correlation-regression analysis, and established its regression equation.

RESULTSThe amount of energy consumption needed in vaporizing one gramme prostatic tissue was (6.9 ± 0.6)kJ. The correlation of energy consumption and preoperative volume of prostate was significantly positive linear correlated. Its regression equation was: Energy Consumption (kJ) = 4.7 x Preoperative Volume of Prostate (cm³) - 14.1.

CONCLUSIONIt is feasible to estimate energy consumption of green laser in PVP based on preoperative volume of prostate.

Aged ; Aged, 80 and over ; Feasibility Studies ; Humans ; Laser Therapy ; Male ; Middle Aged ; Prostate ; pathology ; Prostatic Hyperplasia ; pathology ; surgery ; Transurethral Resection of Prostate ; methods

OBJECTIVEInflammation plays a critical role in secondary brain damage after intracerebral hemorrhage (ICH). However, the mechanisms of inflammatory injury following ICH are still unclear, particularly the involvement of NLRP3 inflammasome, which are crucial to sterile inflammatory responses. In this study, we aim to test the hypothesis that NLRP3 signaling pathway takes a vital position in ICH-induced secondary inflammatory damage and detect the role of N-methyl-D-aspartic acid receptor 1 (NMDAR1) in this progress.

METHODSICH was induced in mice by microinjection of hemin into the striatum. The protein levels of NMDAR1, NMDAR1 phosphorylation, NLRP3 and IL-1b were measured by Western blot. The binding of NMDAR1 to NLRP3 was detected by immunoprecipitation.

RESULTSThe expression of NMDAR1, NMDAR1 phosphorylation, NLRP3 and IL-1b were rapidly increased after ICH. Hemin treatment enhanced NMDAR1 expression and NMDAR1 phosphorylation, as well in cultured microglial cells treated by hemin. Hemin up regulated NLRP3 and IL-1b level, which was reversed by MK801 (NMDAR antagonist) in vitro. Hemin also promoted the binding of NMDAR1 to NLRP3.

CONCLUSIONOur findings suggest that NMDAR1 plays a pivotal role in hemin-induced NLRP3-mediated inflammatory damage through synergistic activation.

Animals ; Cells, Cultured ; Cerebral Hemorrhage ; complications ; Hemin ; pharmacology ; Inflammation ; etiology ; Mice ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein ; physiology ; Phosphorylation ; Receptors, N-Methyl-D-Aspartate ; physiology ; Signal Transduction

Adult ; Anti-Retroviral Agents ; therapeutic use ; DNA, Viral ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Male ; Treatment Outcome ; Viral Load

OBJECTIVETo investigate the effects of cordyceps acid and cordycepin on the inflammatory phenotype and fibrogenic property of hepatic stellate cells (HSCs).

METHODSAn immortalized mouse HSC line (JS1) was stimulated with lippolysaccharide (LPS; 100 ng/ml) to induce an inflammatory response with or without co-administration of cordyceps acid or cordycepin in various concentrations (10, 50, or 200 mumol/L). Effects of the treatments on the chemokine monocyte chemotactic protein-1 (MCP-1) mRNA expression in the cells and the protein secretion in the cell culture supernatants were determined by reverse transcription and real-time quantitative PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. In addition, JS1 cells were treated with transforming growth factor-b1 (TGFb1; 10 ng/ml) to induce a fibrogenic response with or without co-administration of cordyceps acid or cordycepin in various concentrations (10, 50, or 200 mumol/L). Effects on the expression of fibrogenic proteins including collagen type I and a-smooth muscle actin (a-SMA), were investigated by Western blot.

RESULTSHigh-concentration (200 mumol/L) treatments of both cordyceps acid and cordycepin significantly inhibited the LPS-induced up-regulation of MCP-1 transcription and secretion (mRNA: 2.07 +/- 0.29 vs. 3.35 +/- 0.26, t = 15.90 and 1.15 +/- 0.23 vs. 4.17 +/- 0.61, t = 8.93; protein: 1.88 +/- 0.06 vs. 2.33 +/- 0.06, t = 10.39 and 1.47 +/- 0.25 vs. 1.97 +/- 0.04, t = 4.60; all P less than 0.05). All concentrations of cordyceps acid and cordycepin inhibited the TGFb1-induced up-regulation of collagen type I and a-SMA protein expression. However, the effects were more robust with the 200 mumol/L concentrations (P less than 0.05).

CONCLUSIONCordyceps acid and cordycepin ameliorate the LPS-induced inflammatory phenotype and TGFb1-induced fibrogenic response of cultured HSCs. These effects may contribute significantly to the drugs' therapeutic mechanisms to inhibit and resolve liver fibrosis.

Animals ; Cells, Cultured ; Chemokine CCL2 ; metabolism ; Cordyceps ; Hepatic Stellate Cells ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Up-Regulation ; drug effects