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 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.

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.

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.

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.

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 investigate the value of conventional ultrasound combined with contrast‐enhanced ultrasound in qualitative diagnosis of benign and malignant cervical lymph nodes (LNs) .Methods Totally 129 enlarged LNs in 112 cases were performed the 2‐dimensional ,color Doppler ultrasound ,and grey‐scale contrast‐enhanced ultrasound(CEUS)examinations .LNs were divided into the benign group and malignant group according to pathologic results .The aspect ratio ,eccentric medulla or medulla de‐fect ,echo type ,blood supply distribution ,contrast‐enhancement pattern and distribution equilibriuym of contrast agent were com‐pared between the two groups .Results The most of aspect ratio in the benign LNs group were ≥ 2 ,the cortex and medulla were uniform with symmetric increase and without microcalcification ,the blood supple was mainly the non‐blood flow type or hilar type , in contrast‐enhanced ultrasound ,the majority were the early hilar and medullar enhancement ,showing the centrifugally full filling ;the most of aspect ratio in the malignant LNs group were <2 ,medulla was eccentric or loss ,partial of LNs showed the increased parenchymal echo accompanied with cystic lesion and microcalcification ,the blood supply was mainly the peripheral and mixed types ,and the non‐hilar and medulla started enhancement pattern .The ultrasonographic parameters mentioned above were statisti‐cally different between the two groups ,in which the non‐hilar and medulla started enhancement patterns had the highest sensitivity and accuracy for diagnosing malignant LNs ,the positive predictive values of these parameters were higher ,whereas the negative predictive values were lower .Conclusion The conventional ultrasound combined with contrast‐enhanced ultrasonography has a higher practical value in qualitative diagnosis of cervical LNs ;the comprehensive judgment of multiple indicators can increase the differentiating ability for benign from malignant LNs .

AIM To investigate the protective effects of polysaccharides from Dendrobium nobile Lindl.on cerebra ischemia/reperfusion injury in rats and its possible mechanism of action.METHODS One hundred and five SD rats were divided into sham group,model group,Nimodipine group (10 mg/kg) and treatment groups with low-,medium-,and high dose (50,100,and 200 mg/kg) of polysaccharides from D.nobile.The right middle cerebral artery of rats was occluded by inserting a.thread through internal carotid artery for 2 h,and was sampled after reperfusion for 24 h.The rats received gavage once a day for 7 d before operation.Neurological deficits score,brain index,brain water content and infarct size in rats were conducted at the end of reperfusion;the activity of superoxide dismutase (SOD),glutathione peroxidase (GSH-Px),myeloperoxidase (MPO) and the content of malondialdehyde (MDA) of brain tissue and blood serum were measured by chemical colorimetry;the infiltration of neutrophile granulocyte in rat ischemic cortex were detected with immune-fluorescence staining.RESULTS Sham group had no neurological deficit,but the model group showed severe neurological deficits,meanwhile,the infarct size,brain index and brain water content rose markedly,the content of MDA and the activity of MPO of the brain tissue and the blood serum increased remarkably,while the activity of SOD and GSH-Px apparently decreased;compared with the model group,neurological deficits of rats were improved significantly in polysaccharides from D.nobile dose groups,moreover,the infarct size,brain index and brain water content markedly declined,the MDA level and the activity of MPO significantly decreased,but the activity of SOD,GSH-Px increased remarkably;the infiltration of neutrophile granulocyte in the high dose group significantly decreased compared with the model group.CONCLUSION Polysaccharides from D.nobile has some neuroprotective effects on local cerebral ischemia/reperfusion injury,and that may be related to the remove of oxygen free radicals and the decrease in inflammation reaction.

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.

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.