Objective To investigate the regulation mechanism of dendrobium nobile polysaccharides (DNP) for nuclear factor kappa B (NF-κB) signal pathway in focal cerebral ischemia/reperfusion rats.Methods According to the digital random method,90 SD rats were divided into 6 groups:sham operation,model,DNP low-dose (DL,50 mg/kg),moderate-dose (DM,100 mg/kg) and high-dose (DH,200 mg/kg),and nimodipine (10 mg/kg) groups (n =15 in each group),and according to the random number method,selecting 5 in each group for the corresponding index detection.A focal cerebral ischemia/reperfusion model in SD rats was induced by middle cerebral artery occlusion.The improvement effect of DNP on rat neurological deficit (Bederson behavioral score) and brain water content,and infarct volume were observed.Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of proinflammatory cytokine tumor necrosis factor-α (TNF-α) and interleukin-β (IL-1 β) in brain tissue.Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect microglial cell marker BCL-2-related protein A1 α (A1) and astrocyte marker glial fibrillary acidic protein (GFAP) mRNA transcription levels.Western blot was used to detect the NF-κB signaling pathway phosphorylated IκBα protein and nuclear p65 protein expression levels.Single factor variance analysis was used to compare the measurement data among the groups.Results (1) There were significant differences in the neurological deficit score,brain water content,cerebral infarction volume,brain tissue TNF-α and IL-1β levels,A 1 and GFAP mRNA transcription levels and phosphorylated IκBα protein and nuclear p65 protein levels among the 6 groups (F =22.24,8.699,33.89,19.26,27.53,109.5,15.28,66.86,and 41.63,respectively (all P ＜ 0.01).(2) The neurological deficit score,cerebral water content,and cerebral infarction volume in the model group were 2.8 ± 0.3,86.1 ±3.8％,and 31.0 ±4.5％,respectively.The TNF-α and IL-1β levels,A1 and GFAP m RNA transcription levels in brain tissue,and phosphorylated IκBα protein and nuclear p65 protein levels were increased significantly compared with those in the sham operation group.There were significant differences (all P ＜0.01).(3)The above indices in the DH group were 1.5 ± 0.5,72.9 ±5.4％,and 17.5 ±4.1％,respectively.Compared with the model group (including TNF-α and IL-1 β levels,A1 and GFAP mRNA transcription levels and phosphorylation of IκBα protein and nuclear p65 protein levels in brain tissue).There were significant differences (all P ＜ 0.05).Compared with the nimodipine group,there were no significant differences (all P ＞ 0.05).Compared with the sham operation group,except for there were no significant difference in brain water content,phosphorylated IκBα protein and nuclear p65 protein (P ＞0.05),there were significant differences in other indices (all P ＜ 0.05).(4) Compared with the model group,only IL-1 β and phosphorylated IκBα protein levels were decreased significantly in the DM group,there were no significant differences in other indices (all P ＞ 0.05).Compared with the sham operation group,there were significant differences in other indices (all P ＜ 0.05).(5) Compared with the model group,there were no significant differences in other indices in the DL group (P ＞ 0.05).Compared with the sham operation group,there were significant differences in all indices (all P ＜ 0.05).Conclusion Highdose DNP may reduce focal cerebral ischemia/reperfusion injury in rats.The mechanism of may be associated with the inhibition of activation of early NF-κB signaling pathway.The effects of low-and moderate-dose DNP on reducing inflammatory brain damage of focal cerebral ischemia/reperfusion may be not obvious.

Objective:To study the transdermal absorption in vitro of sinapine thiocyanate,tetrahydropalmatine and asarinin in Xiaochuan ointment so as to offer reference for its clinical application.Methods:A Franz diffusion cells method with isolated rat skins was carried out to study the percutaneous rate of sinapine thiocyanate,tetrahydropalmatine and asarinin determined by LC-MS/MS.Results:With the increase of administration dosage,the cumulative penetration of sinapine thiocyanate,tetrahydropalmatine and asarinin showed few changes.The results showed that the transdermal behavior of sinapine thiocyanate fit to a Higuchi equation,and that of tetrahydropalmatine and asarinin fit zero-order process.The penetration rate of tetrahydropalmatine and asarinin respectively was 0.362×10-1 and 0.330×10-2 μg·cm-2·h-1.Conclusion:Xiaochuan ointment exhibits transdermal penetration and absorption properties,which provides evidence for its transdermal administration.

In this paper, the morphological development of periderm of the hands and feet of 21 human embryos were investigated. The estimated gestational age (EGA) of the collected specimens were 6-24 weeks.On 36th to 41st days of EGA, the cells of embryonic periderm were seen to be flat and polygonal in shape. Microvilli were seen on the cell surface. At the end of 7th week the cells showed conspicuous central elevations as cobblestones. After 11th week, a large single bleb occurring on the cell surface were observed. At 4th month, the number of the blebs increased and complex blebs were formed. Larger blebs showed infoldings and furrows. Then the blebs disappeared and the cell surface became flat again. The cells of periderm began to shed at 17th week.The development of periderm on the hands appeared earlier than that on the feet. The degree of the development of periderm in different regions of hands and feet may be different. The possible significance of the periderm was discussed.

Objective To investigate the effects of recombinant human pigment epithelium derived factor (rhPEDF)on in vitro proliferation of and expressions of interleukin 6(IL-6), IL-8 and vascular endothelial growth factor (VEGF)in cultured human HaCaT keratinocytes. Methods Some cultured HaCaT cells were treated with rhPEDF at various concentrations(25, 50, 100 μg/L)for different durations, and some treated with RPMI 1640 medium only served as the control group. Cell counting kit-8(CCK8)assay was performed to evaluate cell proliferation after 24-, 48- and 72-hour treatment, reverse transcription (RT)-PCR to measure the mRNA expressions of IL-6, IL-8 and VEGF in HaCaT cells after 24-hour treatment, and Western blot to detect the protein expressions of IL-6, IL-8 and VEGF in HaCaT cells after 48-hour treatment. Statistical analysis was carried out by two- and one-way analysis of variance, Student-Newman-Keuls(SNK)-q test and Pearson correlation analysis. Results After treatment with rhPEDF of 25-100 μg/L for 24 - 72 hours, the proliferation of HaCaT cells was significantly inhibited to different extents compared with the control group(all P < 0.05), and the inhibition rate significantly increased with the increase in treatment duration and concentrations of rhPEDF(F = 1115, 329.9, respectively, both P < 0.001). Moreover, there was a significant decrease in the expressions of IL-6, IL-8 and VEGF mRNAs(at 24 hours)and proteins(at 48 hours)in HaCaT cells after treatment with rhPEDF of 25 - 100 μg/L compared with the control group(all P < 0.05). The expression levels of VEGF mRNA as well as IL-6 and IL-8 proteins all significantly decreased with the increase of rhPEDF concentrations (all P < 0.05). The mRNA expressions of IL-6 and IL-8 were significantly lower in the 100-μg/L rhPEDF group than in the 25-μg/L rhPEDF group (both P < 0.05), and the protein expression of VEGF was significantly weaker in the 100-μg/L rhPEDF group than in 25-and 50-μg/L rhPEDF groups (both P < 0.05), but similar between the 25- and 50-μg/L rhPEDF groups (P > 0.05). Conclusions rhPEDF can inhibit the proliferation of HaCaT cells, and down-regulate the mRNA and protein expressions of IL-6, IL-8 and VEGF.

Objective To investigate the diagnosis and treatment of post-transplant lymphoproliferative disease (PTLD) after hematopoietic stem cell transplantation (HSCT).Methods Clinical data of one case of PTLD,including clinical manifestations,diagnosis and treatment,were retrospectively analyzed.And the related literatures were reviewed.Results From January 2007 to November 2012,one case developed PTLD among 196 patients who were followed up after HSCT in the First Affiliated Hospital of Chongqing Medical University.The incidence of PTLD was 0.5％.Clinical manifestations of the patient were not typical,including frequent fever,multiple lymphadenopathy and multiple pulmonary nodules.The patient was diagnosed as diffuse large B-cell lymphoma (B-DLCL) by pathological examination of the swollen lymph node.After withdrawal of immunosuppressants,the temperature returned to normal,and lymphadenopathy and pulmonary nodules disappeared completely.Conclusion PTLD is a severe complication of HSCT with distinctive morphologic and clinical characteristics.The origin of this disease may be associated with Epstein-Barr virus infection and imrnunosuppressive therapy.It needs the pathologic detection to make a definite diagnosis.Several different treatment strategies have been employed,and reduction of immunosuppressive therapy may lead to regression of PTLD.

AIM: To study the biology characteristics of mesenthymal stem cells(MSCs) derived from chronic myelogenous leukemia(CML) and normal adult bone marrow.METHODS: Mononuclear cells from chronic myelogenous leukemia(n=19) and normal adult(n=8) bone marrow were obtained,cultured in expanded medium with low serum concentration.Cell morphology,cell cycle,immunophenotype and in vitro differentiation capacity were investigated.The differentiations of osteocytes and adipocytes were detected by von Kossa staining and Oilred O staining.The chimeric oncogene BCR/ABL and Ph chromosome,two hall marks of CML,were detected in CML derived MSCs,normal adult MSCs,CML derived hematopoietic cells and K562 cells.RESULTS: CML and normal adult derived MSCs showed similar characteristics in cell morphology,phenotype and growth pattern.A typital fibrablast like morphology was observed.Under suitable conditions,CML and normal adult MSCs had the similar ability to differentiate into adipocytes and osteoblasts in vitro.Moreover,CML and normal adult MSCs did not express BCR/ABL gene products and Ph chromosome was not observed.CONCLUSIONS: We isolated and cultured a population of cells with characteristics of multipotent stem cells from CML bone marrow.There were similar biologic characteristics and differentiation ability between normal adult and CML bone marrow-derived MSCs.

AIM: To investigate the isolation,purification,expansion and multilineage differentiation of mesenchymal stem cells(MSCs) derived from human umbilical cord vein in vitro.METHODS: By 1% collagenase Ⅱ digestion,endothelial cells were isolated from human umbilical cord vein and cultured in IMDM medium.The morphology of the cells was observed by Wright's staining and electron microscope.Cell cycle and immunophenotype were investigated by flow cytometry.Assays of adipogenic and osteogenic differentiation were performed in vitro.von Kossa staining,Oil Red O staining and mRNA expression of osteopontin and lipoprotein lipase were studied in the induced cells.RESULTS: The cells from the cord vein displayed a fibroblast-like morphology adhering to the culture plate.FACS showed that the cells expressed several MSCs-related antigens such as CD29,CD44 and CD105,while CD13,CD31,CD45,CD34,and HLA-DR were negative.Adipocyte and osteocyte differentiation were induced successfully.CONCLUSION: The morphology,growth characteristics,immunophenotype and pluripotentiality of the MSCs from human umbilical cord vein are similar to the MSCs from bone marrow(BM).They could potentially be an excellent source of MSCs for experiments and clinics.

The surface morphology and histogenesis of small intestine of human embryos (from 6 to 36 weeks) were studied by scanning electron microscopy and light microscopy. The results indicated that some low prominences began to form at 6 weeks of gestation and then transformed to round tuberculum, after that to polygonal column. The finger-like intestinal villi were formed finally at the end of 3 months. There was a apparent proximodistal gradient of the morphological maturation of villi and the histogenesis of intestine from duodenum, jejunum to ileum. Splitted and branched villi were found. From 18 weeks onward, the respective characteristics of villi in the three segments of small intestine appeared. Neutral mucus and sialomucus were mainly secreted by the goblet cells, while sulphomucus was secreted by the goblet cells of ileum during the late gestation. By 6-12 weeks the microvilli of duodenum were shorter than that of jejunum and ileum but they increased in height to 1.4?m at 18 weeks. The height of microvill at jejunum and ileum were 0.8-1.0?m from 14 weeks to 32 weeks. Tufts of thick and long microvilli among short microvilli were found. The formation of villi, secretion of goblet cells and microvilli was discussed.

The immune mechanism of Graves' diseases (GD) and the roles of regulator T cells were investigated. In 32 patients with GD (GD group) and 20 healthy volunteers (control group), flow cytometry was used to detect the proportion of CD4+CD25+ cells, MACS to isolate CD4+ CD25+ cells, RT-PCR to assay the expression of FOXP3, and ELISA to test the level of IL-10, respectively. It was found that there was no significant change in the proportion of CD4+CD25+ T cells between GD group and control group (P>0.05), while secretion of IL-10 and expression of FOXP3 in GD group were lower than control group (P<0.01 and P<0.05, respectively). In conclusion, though the proportion of regulatory T cells of peripheral blood lymphocytes in the patients with GD, the functions of them were significantly weakened, which might be a pathogenic factor in GD.

Objective To study the correlation between hyperhomocystinemia and the property of cerebral artery plaque,and the pathogenic mechanism of hyperhomocystinemia resulting in acute cerebral stroke.Methods 60 patients with acute ischemic stroke were chosen.30 patients with normal homocysteine were classified as control group.The other 30 patients with hyperhomocystinemia were classified as observation group.High resolution magnetic resonance imaging(HRMRI) and multi contrast plaque imaging technique were used to measure the cerebral artery plaque burden,the fat ratio of plaques,the hemorrhage within the plaque and fibrouscap rupture.The number of inflammatory cells in the plaque and the density of the new blood vessels were identified using the transfer constant of the contrast medium.Results In the observation group,4 cases with low-risk plaques (13.33％),10 cases with middle-risk plaques (33.33 ％) and 16 cases with high-risk plaques (53.33 ％).In the control group,13 cases with low-risk plaques (43.33％),6 cases with middle-risk plaques (20.00％),11 cases with high-risk plaques (36.67％).The number of intraplaque hemorrhage were 18 cases (60.00％) in the observation group,and 8 cases (26.67％) in the control group.The number of fibrouscap rupture were 21 cases (70.00％) in the observation group,and 13 cases (43.33％) in the control group.Risk stratification of vulnerable plaques in cerebral atherosclerosis (U =-2.032,P =0.042),intraplaque hemorrhage (x2 =6.79,P =0.009) and fibrouscap rupture (x2 =4.34,P =0.037) in the observation group was higher than that in the control group.The number of inflammatory cells in plaques and the density of the new blood vessels in the observation group was (0.188 ± 0.265)Ktrans/min,which in the control group was (0.118 ± 0.183)Ktrans/min.The number of inflammatory cells in the plaque and the density of the new blood vessels in the observation group was higher than that in the control group (t =11.831,P =0.000).Conclusion Hyperhomocystinemia causes intraplaque hemorrhage,fibrouscap rupture,inflammatory cell activity and neovascularization in plaques,accelerates cerebral atherosclerosis and increases the incidence of ischemic stroke events.