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.

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

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.

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.

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

Objective:To obtain the inter-fractional set-up errors of intensity-modulated radiotherapy (IMRT) of cervical cancer by cone-beam CT (CBCT), and to analyze the variations of the set-up errors on the cumulative dose deviation of the target volume.Methods:A total of 48 patients with cervical cancer who underwent IMRT were enrolled in this study, and the set-up errors of 696 CBCTs were obtained. The set-up errors were input into the treatment planning system, and the cumulative set-up error dose was obtained by superposing the set-up errors dose. The deviation percentage was calculated by the deviation formula and the standard planning dose.Results:The set-up errors caused the offset of isocenter distance by 0.58(0.36, 0.80) cm. Different statistical differences were noted between the cumulative set-up error dose and the standard planning dose by WilCoxon test. All the dose deviations in the target volume were reduced, and the differential dose volume histogram (DVH) appeared negatively skewed, and the peak value was declined. The DVH diagram shifted to the left with an inverse S-curve and the slope was increased. The HI deviation of the target volume from small to large were: CTV 1, CTV 2, GTV/CTV, CTV 3, CTV n, CTV all, and GTV nd; The HI deviation of the target volume were increased. Conclusions:The effect of set-up errors in IMRT of cervical cancer upon the cumulative doses of the target volume significantly differs. The cumulative dose of the target volume is reduced, and the uniformity of the target volume dose becomes worse. The uncertainty of the inter-fractional position leads to an increase or decrease in the the fractional doses of the target volume. The biological effect on tumor cells and the tumor recurrence remains to be investigated. In IMRT of cervical cancer, the CBCT position calibration is required before each treatment to ensure the dose accuracy of each structure in the target volume.

Objective: To investigate the genetic characteristics of Lamivudine-resistant mutation patterns and HBV S gene mutants in patients with chronic hepatitis disease of different disease progression. Methods: Blood samples of LAM-resistant patients with chronic hepatitis disease were collected. HBV RT gene nucleotide sequences were obtained, and then differences in drug-resistant mutation patterns, drug susceptibility and HBV S gene mutants characteristics between the two groups were analyzed. Results: Forty-seven chronic hepatitis B (CHB) patients and 16 HBV-related liver cirrhosis (LC)/HBV-related hepatocellular carcinoma (HCC) patients were included in this study. M204I single point mutation and L180M+ M204I/V were the most common pattern during patients with chronic hepatitis disease (35/63, 55.56%). The numbers of resistant to three nucleos(t)ide analogues in LC/HCC group was higher than CHB group’s (62.50% vs 34.04%, P=0.046). In HBV S gene, more immune associated HBsAg-escape mutations were detected in LC/HCC group than that in CHB group (62.50% vs 31.91%, P=0.031). I126T/V and G145A (for LCC/HCC group, 60%), I126S/T and S117T (for CHB group, 46.67%) were showed as the most common form for HBsAg escape mutations in the two groups. The two groups both detected RT mutations concomitantly with stop codon mutations in S gene (rtA181T/sW172* and rtM204I/sW196*). Conclusions Different characteristics in Lamivudine-resistant mutations and associated HBV S gene mutants were found in patients with chronic hepatitis disease of different disease progression, and LC/HCC patients exhibit more multi-drug resistant variants and immune associated HBsAg-escape mutants than CHB patients.

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

OBJECTIVETo make an attempt at the multi-element speciation in the Chinese medicinal herbs by determining the concentrations of 25 elements in different extraction solutions.

METHODFirstly, five Chinese medicinal herbs (Buddleja officinalis, Dictamnus dasycarpus, Myristica fragrans, Albizia judibrissin and Inula japonica) from the same region of China were treated to obtain water-soluble phase, lipid-soluble phase and non-soluble phase by water extraction, organic solvent extraction and acid digestion, respectively. Secondly, Phytolacca acinosa, a Chinese medicinal herb collected from 9 regions of China, was extracted by 0% EtOH, 50% EtOH, 75% EtOH, 95% EtOH, respectively, referring the Chinese Pharmacopoeia. Finally, the concentrations of 25 elements, such as Be, Cr, Cu, Zn, Ge, Sr, Y, Mo, Cd, Tl, Pb and REEs, in the above three phases were determined by ICP-MS.

RESULTUnder the optimal conditions, all the 25 elements could be determined with detection limits ranged from 0.003 to 0.71 ng x g(-1). The average recoveries of the elements in P. acinosa were 88% approximately 119%, with the relative standard deviations 1.7% approximately 13.3%. It was observed that the determined 25 elements distributed in all the water-soluble, lipid-soluble and non-soluble phases, indicating that the inorganic species, organicspecies, as well as the protein bound species were coexisted in the herbs. Big differences of the element extraction rates could be found by using different ethanol solutions.

CONCLUSIONWith the aid of the obtained results, we may increase the extraction of necessary elements while decrease that of the toxic elements from the herbs by choosing a suitable solvent during the drug production.

Buddleja ; chemistry ; Cadmium ; analysis ; Copper ; analysis ; Dictamnus ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ecosystem ; Lead ; analysis ; Metals, Heavy ; analysis ; Molybdenum ; analysis ; Myristica fragrans ; chemistry ; Phytolacca ; chemistry ; Plants, Medicinal ; chemistry ; Solvents ; chemistry ; Trace Elements ; analysis ; Zinc ; analysis