TLC method for the identification of triterpenoids polyphenols in Fructus Chebulae was established .The results showed that this method had a plentiful chromatogram and good specificity,and can distinguish Fructus Chebulae immaturus from Fructus Chebulae.Determination of gallic acid, one of the hydrolysates of Fructus Chebulae, by HPLC was also reported.The linear range of gallic acid is from 0.17?g to 1.36?g(r=0.9998),and the average recovery is 102.9 %. The results showed that the method is simple, accurate and with good reproducibility.It can be used to determine gallic acid in Chinese herbal medicine that contains hydrolysable tannins.

Aim A method of coculture of brain capillary endothelial cells (BCECs) and astrocytes of rats was used to evaluate nanoparticle' s blood-brain barrier (BBB) transcytosis and toxicity at the endothelial tight junction. Methods A lipophilic fluorescent probe, 6-coumarin, was incorporated in poly(ethyleneglycol)-poly (lactide) nanoparticle using double emulsion/solvent evaporation method. BCECs and astrocytes were firstly isolated from brain of newborn rats and characterized by their morphology and immunocytochemistry staining, separately. Subsequently, a coculture model with BCECs on the top of micro-porous membrane of cell culture insert and astrocytes on the bottom side was established. The permeability of 14C-labeled sucrose and nanoparticle were determined, separately. Results The meanweight-based diameter of 6-coumarin loaded nanoparticles was ( 102.4 ± 6.8) nm, with zeta potential of ( - 16.81 ± 1.05) mV. BCECs were positive for factor Ⅷ staining and glial fibrillary acidic protein was tight junction between BCECs in the coculture model could be visualized by both scanning electron microscopy and transmission electron microscopy. The unchanged paracellular transport of sucrose proved vivo situation for examination of the permeability of nanoparticle and toxicity evaluation.

Objective Stem cell transplantation is a new approach to the treatment of lower limb ischemia ( LLI) .This study was to investigate the therapeutic effect of the transplantation of autologous bone marrow mesenchymal stem cells ( BM-MSCs) in the treatment of LLI. Methods We established the left LLI model in 12 New Zealand rabbits and divided them into a control and a trial group of equal number.The control animals were injected with DMEM, while the rabbits in the trial group with autologous BM-MSCs, into the ischemic skeletal muscle.Four weeks after injection, we performed CT angiography and perfusion imaging of both lower limbs of the rabbits and conducted HE staining of the paraffin sections of the skeletal muscle of the ischemic limbs. Results Dynamic ob-servation revealed different degrees of ecderon necrosis in 2 of the LLI models in the control group, even with toenail coloboma, but no necrosis in the trial group except for some slight muscular atrophy. Both collateral arteries and blood perfusion were obviously increased in the ischemic lower limbs in the trial than in the control group.HE stai-ning showed a significantly higher density and percentage of capillaries in the skeletal muscle fibers in the former than in the latter ([6.500 ± 1.049]/HP vs [3.670 ±0.816]/HP, [9.68 ±0.56]%vs [5.87 ± 0.86]%, P<0.01), with no necrosis in either group, nor hematoma, bony tissue, or fibroid tumor in the trial group. Conclusion Autologous transplantation of BM-MSCs can improve neovascularization in ischemic lower limbs in rabbits and can be used as a safe and effective treatment of limb ischemia.

As seculars,we often feel unhappy,and it seems all of our unhappiness comes from almost every aspect of life,which is unable to escape.Russell′s view of happiness dug out the root of unhappiness from human nature,and tells us happiness could be obtained by expending one′s interests and necessarily giving away something in life.Russell′s great wisdom opens a door for us to the road of happiness.

Objective To study the effects of Ganoderma lucidum polysaccharide (GLP) on PBMCs and the related immune mechanism.Methods PBMCs from cancer patients and healthy donors were isolated and treated with different doses of Ganoderma lucidum polysaccharides (10 ng/ml,50 ng/ml and 100 ng/ml).DC cell costimulatory molecules (HLA-DR,CD83,and CD11 c),Th1 (CD3 + CD8-IFN-γ+) cells,Th2 (CD3 + CD8-IL-4+) cells and NK (CD3-CD56+) were analyzed by FCM.Furthermore,The CD3+ CD4+ Th ceils were separated by immunomagnetic beads and stimulated with Ganoderma lucidum polysaccharides at different concentrations in culture.After 24 h,the cytokine expression levels of Th1 and Th2were detected by RT-PCR.The expressions of Th1 differentiation-related transcription factor,STAT4,were analyzed by Western blot.Results Ganoderma lucidum polysaccharides can significantly stimulate in vitro Thl cell differentiation (P＜0.01) in a dose depend manner.It correlates with an increased expression of STAT4 and the elevated mRNA expression levels of Th1 cytokineincluding IL-12,IFN-γ and TNF-α (P＜0.01).Conclusion Ganoderma lucidum polysaccharides may promote Th1 differentiation and increase the secretion of Th1 cvtokines through the upregulation of STAT4.

OBJECTIVE:To observe the effects of different routes of administration of tranexamic acid on coagulation function and amount of bleeding in patients with one stage posterior surgery of thoracic tuberculosis. METHODS:40 patients suffered from thoracic tuberculosis in our hospital from Jan. 2011 to Dec. 2013 were randomly divided into intravenous group(5% Glucose injec-tion 100 ml+tranexamic acid 10 mg/kg,through an intravenous drip at 30 min before closing the wound) and topical application group(5% Glucose injection 10 ml and tranexamic acid 10 mg/kg,through soaking the wound before closing the wound)with 20 cases in each group. Other 15 cases suffered from the thoracic tuberculosis in our hospital from Jan. 2009 to Dec. 2010 were includ-ed in control group. 3 groups received one stage posterior surgery of thoracic tuberculosis,interbody fusion and internal fixation. The difference of hemoglobin,coagulation function and the amount of suction drainage were observed before and after surgery, and followed up. Bone graft fusion and therapeutic condition of tuberculosis were observed in the study. RESULTS:There was no statistical significance in postoperative suction drainage between intravenous group and topical application group (P>0.05),but their decrease was more significant than control group,with statistical significance(P<0.05). There was no statistically difference in fiber protease,prothrombin time or activated partial thromboplastin time among 3 groups(P>0.05). The difference value of he-moglobin in control group before and after operation was significantly higher than in intravenous group and topical application group,with statistical significance (P<0.05);there was no statistical significance between intravenous group and topical applica-tion group(P>0.05). 55 patients were all followed up and bone graft of all cases were fused,and all patients were cured and no case recurred. CONCLUSIONS:Tranexamic acid by intravenous application or topical application can reduce hemorrhage and ane-mia after operation of thoracic tuberculosis,and has no effect on blood coagulative system.

Objective To observe the effects of mesenteric lymph drainage on acute lung injury and expression of p38 mitogen activated protein kinase (p38MAPK) signal pathway in rats with bowel repletion pattern. Methods Thirty male Wistar rats were randomly divided into three groups according to random number table method, namely sham operation group (sham group), bowel repletion model group (model group) and mesenteric lymph drainage group (drainage group), 10 rats in each group. The rat model of bowel repletion was established by ischemia/reperfusion (I/R) method, firstly 1 hour occlusion of superior mesenteric artery (SMA) to induce ischemia followed by reperfusion for 2 hours. In the rats of drainage group, the drainage of mesenteric lymph duct began at the end of model establishment and persisted for 3 hours. In the rats of sham group, the SMA and mesenteric lymph ducts were exposed with blunt dissection, and then they were immediately placed back into the abdominal cavity. After 3 hours of mesenteric lymph drainage, the lung and ileum tissues of rats in each group were harvested for evaluation of pathohistological changes and for the determination and comparison of myeloperoxidase (MPO) activity changes; the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay (ELISA), and the expressions of Toll-like receptor 4 (TLR4) mRNA and p38MAPK mRNA in the lung tissues were measured by fluorescence quantitative polymerase chain reaction (PCR).Results Under the light microscope, the pulmonary capillaries markedly dilated and congested, the interstitium width of lung increased with a large amount of inflammatory cells infiltration, the intestinal mucosal layer becoming thinner with detachment of intestinal villi and a large amount of inflammatory cells infiltration were detected in rats of model group. Compared with those in sham group, the levels of TNF-α and IL-6 in BALF, the MPO activity of lung and ileum tissues, and the expressions of TLR4 mRNA and p38MAPK mRNA in the lung tissues were significantly increased in model group.Compared with those in model group, the pathohistological damages in lung and ileum tissues were ameliorated, the levels of TNF-α and IL-6 in BALF, the MPO activity of lung and intestinal tissues and the expressions of TLR4 mRNA and p38MAPK mRNA in the lung tissues were lower in the rats of drainage group [TNF-α in BALF (ng/L): 858.55±27.16 vs. 1 680.58±105.62; IL-6 in BALF (ng/L): 0 vs. 484.71±5.43; MPO activity of lung (U/g): 0.95±0.13 vs. 1.36±0.11; MPO activity of ileum tissues (U/g): 0.75±0.13 vs. 1.30±0.16; TLR4 mRNA: 0.21±0.11 vs. 0.69±0.13, p38MAPK mRNA: 0.21±0.13 vs. 0.47±0.09; allP < 0.05].Conclusion Mesenteric lymph drainage can alleviate acute lung injury in rats with bowel repletion, and its mechanism may be related to the reduction of the expressions of TLR4 mRNA and p38MAPK mRNA and the release of TNF-α and IL-6 in lung tissues.

Soil particles are very heterogeneous in microscopic scale, which is manifested the double-layer structure made of the soil organic matter and mineral matter. In this work, Fourier by transform infrared photoacoustic spectroscopy ( FTIR-PAS) combined with independent component analysis ( ICA) was utilized for in situ depth-profiling of the manmade complex film, in order to lay a foundation of in situ characterizing the heterogeneous soil organic-mineral complex. The complex film was composed of the PE preservative film and office adhesive tape. The moving velocity of infrared photoacoustic spectrometer was set to 0. 16 cm/s, 0. 32 cm/s and 0. 64 cm/s, respectively. Independent component analysis ( ICA ) was performed on the photoacoustic spectra of the heterogeneous complex film. Results showed that the depth-resolved information of the complex film could be derived by changing the moving velocity, and the estimated thickness of PE film was 5. 4-7. 6 μm, which was close to the actual thickness 7 ± 1 μm. Moverover, the spectral features of the polyethylene ( PE) preservative film and office adhesive tape were extracted from the photoacoustic spectra of the heterogeneous complex film by means of ICA. Depth profiling of complex film samples showed that FTIR-PAS could be used as a new analytical tool to study heterogeneous soils, especially soil organic-mineral complexes.

BACKGROUND:Studies have indicated that melatonin can promote the differentiation of adipose-derived stem cels into neurons, and the effect of melatonin on the osteoblasts form adipose-derived stem cels is rarely reported.
OBJECTIVE:To observe the osteogenic differentiation of adipose-derived stem cels and the effect of melatonin on the bio-viability of differentiated cels.
METHODS:(1) Adipose-derived stem cels were isolated and purified from the inguinal fat of Kunming mice by type I colagenase digestion and differential adhesion method, respectively. Immunohistochemical staining of CD44 was used as a quality control. (2) Osteogenic induction medium was added to induce osteogenic differentiation of passage 2 adipose-derived stem cels. Alkaline phosphatase staining and von Kossa method were combined to evaluate differentiation condition. (3) Melatonin at variable concentrations was added to treat mature osteocytes originated from adipose-derived stem cels and MTT was applied to determine their viability at 24 and 48 hours after culture respectively to find out optimal condition of melatonin treatment. (4) Melatonin at the optimal concentration was used to treat differentiated cels and detect alkaline phosphatase activity after 3 days and 6 days respectively.
RESULTS AND CONCLUSION: (1) After seeding for 48 hours, most cels were adherent, and after 4 days, the cels displayed multiple shapes and colonies of different sizes formed. After subculture, cel morphology homogenized as spindle shape. Cels positive for CD44 were brownish yelow, and localized mainly on the cel membrane. (2) Differentiated cels were positive for von Kossa staining and black sediments scattered in the extracelular matrix. Alkaline phosphatase expressed positively, and brown-black particles, appeared within cels. (3) Melatonin supplement improved the viability of differentiated cels; and 1, 10 and 100 μmol/L was observed as the optimal concentrations both at 24 and 48 hours. (4) The intracelular alkaline phosphatatse activity was increased with time in al the groups (P < 0.05). Compared with the blank group, the intracelular alkaline phosphatase activity in Melatonin groups (1, 10 and 100 μmol/L) had nochanges at 3 days, but significantly increased at 6 days (P < 0.05). These findings indicate that melatonin can enhance the proliferation of osteocytes differentiated from adipose-derived stem cels, and improve the activity of intracelular alkaline phosphatase.

Objective Polyinosinic-polycytidylic acid (poly IC) plays an important role in the central nervous system damage and repair.This study was to investigate the effect of poly IC on inflammatory response after spinal cord ischemia-reperfusion injury (SCIRI) in rats.Methods A total of 72 healthy adult male SD rats were equally randomized into a sham-operation, an ischemia-reperfusion (IR), and a poly IC group.The abdominal cavities of the rats were cut open and closed again in the sham-operation group and SCIRI models were established in the IR and poly IC groups by clamping the abdominal aorta, followed by reperfusion 60 minutes later and intraperitoneal injection of saline (0.1 mL) and poly IC (1.25 μg/g), respectively.At 6, 24, and 48 hours after modeling, BBB scores were obtained and the contents of TNFα, IL-1β and IFN-β were measured by ELISA.At 48 hours, the expressions of NF-κB and IL-10 were determined by immunohistochemistry, the area of ischemic necrosis in the spinal cord tissue was calculated by TTC staining, and its morphological changes were observed under the optical microscope.Results At 48 hours after modeling, the BBB scores were significantly lower in the IR and poly IC groups than in the sham-operation group (3.80±0.75 and 9.40±0.49 vs 20.00±0.00, P<0.01), though higher in the poly IC than in the IR group (P<0.01).The rats of the IR group showed extensive degenerated neurons in the gray substance of the spinal cord, with scattered foci of bleeding and blood coagulation, while those of the poly IC group exhibited fewer necrotic neurons and basically normal nuclear morphology, though with a few swelling cells.The ischemic necrosis area of the spinal cord tissue was significantly reduced.The expression of NF-κB was decreased while that of IL-10 increased markedly.Compared with the IR group, the poly IC group showed a significant increase in the expression of IFNβ (117.23±6.06 vs 55.65±4.02, P<0.01) and a remarkably decrease in the expressions of TNFα (190.45±4.16 vs 201.82±2.18, P<0.01) and IL-1β (39.27±2.48 vs 50.59±1.47, P<0.01) at 48 hours.Conclusion Poly IC can protect the spinal cord and reduce inflammatory response after spinal cord ischemia-reperfusion injury.